Isolated human kinase proteins, nucleic acid molecules encoding human kinase proteins, and uses thereof

ABSTRACT

The present invention provides amino acid sequences of peptides that are encoded by genes within the human genome, the kinase peptides of the present invention. The present invention specifically provides isolated peptide and nucleic acid molecules, methods of identifying orthologs and paralogs of the kinase peptides, and methods of identifying modulators of the kinase peptides.

FIELD OF THE INVENTION

[0001] The present invention is in the field of kinase proteins that arerelated to the serine/threonine protein kinase subfamily, recombinantDNA molecules, and protein production. The present inventionspecifically provides novel peptides and proteins that effect proteinphosphorylation and nucleic acid molecules encoding such peptide andprotein molecules, all of which are useful in the development of humantherapeutics and diagnostic compositions and methods.

BACKGROUND OF THE INVENTION

[0002] Protein Kinases

[0003] Kinases regulate many different cell proliferation,differentiation, and signaling processes by adding phosphate groups toproteins. Uncontrolled signaling has been implicated in a variety ofdisease conditions including inflammation, cancer, arteriosclerosis, andpsoriasis. Reversible protein phosphorylation is the main strategy forcontrolling activities of eukaryotic cells. It is estimated that morethan 1000 of the 10,000 proteins active in a typical mammalian cell arephosphorylated. The high-energy phosphate, which drives activation, isgenerally transferred from adenosine triphosphate molecules (ATP) to aparticular protein by protein kinases and removed from that protein byprotein phosphatases. Phosphorylation occurs in response toextracellular signals (hormones, neurotransmitters, growth anddifferentiation factors, etc), cell cycle checkpoints, and environmentalor nutritional stresses and is roughly analogous to turning on amolecular switch. When the switch goes on, the appropriate proteinkinase activates a metabolic enzyme, regulatory protein, receptor,cytoskeletal protein, ion channel or pump, or transcription factor.

[0004] The kinases comprise the largest known protein group, asuperfamily of enzymes with widely varied functions and specificities.They are usually named after their substrate, their regulatorymolecules, or some aspect of a mutant phenotype. With regard tosubstrates, the protein kinases may be roughly divided into two groups;those that phosphorylate tyrosine residues (protein tyrosine kinases,PTK) and those that phosphorylate serine or threonine residues(serine/threonine kinases, STK). A few protein kinases have dualspecificity and phosphorylate threonine and tyrosine residues. Almostall kinases contain a similar 250-300 amino acid catalytic domain. TheN-terminal domain, which contains subdomains I-IV, generally folds intoa two-lobed structure, which binds and orients the ATP (or GTP) donormolecule. The larger C terminal lobe, which contains subdomains VI A-XI,binds the protein substrate and carries out the transfer of the gammaphosphate from ATP to the hydroxyl group of a serine, threonine, ortyrosine residue. Subdomain V spans the two lobes.

[0005] The kinases may be categorized into families by the differentamino acid sequences (generally between 5 and 100 residues) located oneither side of, or inserted into loops of, the kinase domain. Theseadded amino acid sequences allow the regulation of each kinase as itrecognizes and interacts with its target protein. The primary structureof the kinase domains is conserved and can be further subdivided into 11subdomains. Each of the 11 subdomains contains specific residues andmotifs or patterns of amino acids that are characteristic of thatsubdomain and are highly conserved (Hardie, G. and Hanks, S. (1995) TheProtein Kinase Facts Books, Vol I:7-20 Academic Press, San Diego,Calif.).

[0006] The second messenger dependent protein kinases primarily mediatethe effects of second messengers such as cyclic AMP (cAMP), cyclic GMP,inositol triphosphate, phosphatidylinositol, 3,4,5-triphosphate,cyclic-ADPribose, arachidonic acid, diacylglycerol andcalcium-calmodulin. The cyclic-AMP dependent protein kinases (PKA) areimportant members of the STK family. Cyclic-AMP is an intracellularmediator of hormone action in all prokaryotic and animal cells that havebeen studied. Such hormone-induced cellular responses include thyroidhormone secretion, cortisol secretion, progesterone secretion, glycogenbreakdown, bone resorption, and regulation of heart rate and force ofheart muscle contraction. PKA is found in all animal cells and isthought to account for the effects of cyclic-AMP in most of these cells.Altered PKA expression is implicated in a variety of disorders anddiseases including cancer, thyroid disorders, diabetes, atherosclerosis,and cardiovascular disease (Isselbacher, K. J. et al. (1994) Harrison'sPrinciples of Internal Medicine, McGraw-Hill, New York, N.Y., pp.416-431, 1887).

[0007] Calcium-calmodulin (CaM) dependent protein kinases are alsomembers of STK family. Calmodulin is a calcium receptor that mediatesmany calcium regulated processes by binding to target proteins inresponse to the binding of calcium. The principle target protein inthese processes is CaM dependent protein kinases. CaM-kinases areinvolved in regulation of smooth muscle contraction (MLC kinase),glycogen breakdown (phosphorylase kinase), and neurotransmission (CaMkinase I and CaM kinase II). CaM kinase I phosphorylates a variety ofsubstrates including the neurotransmitter related proteins synapsin Iand II, the gene transcription regulator, CREB, and the cystic fibrosisconductance regulator protein, CFTR (Haribabu, B. et al. (1995) EMBOJournal 14:3679-86). CaM II kinase also phosphorylates synapsin atdifferent sites, and controls the synthesis of catecholamines in thebrain through phosphorylation and activation of tyrosine hydroxylase.Many of the CaM kinases are activated by phosphorylation in addition tobinding to CaM. The kinase may autophosphorylate itself, or bephosphorylated by another kinase as part of a “kinase cascade”.

[0008] Another ligand-activated protein kinase is 5′-AMP-activatedprotein kinase (AMPK) (Gao, G. et al. (1996) J. Biol. Chem. 15:8675-81).Mammalian AMPK is a regulator of fatty acid and sterol synthesis throughphosphorylation of the enzymes acetyl-CoA carboxylase andhydroxymethylglutaryl-CoA reductase and mediates responses of thesepathways to cellular stresses such as heat shock and depletion ofglucose and ATP. AMPK is a heterotrimeric complex comprised of acatalytic alpha subunit and two non-catalytic beta and gamma subunitsthat are believed to regulate the activity of the alpha subunit.Subunits of AMPK have a much wider distribution in non-lipogenic tissuessuch as brain, heart, spleen, and lung than expected. This distributionsuggests that its role may extend beyond regulation of lipid metabolismalone.

[0009] The mitogen-activated protein kinases (MAP) are also members ofthe STK family. MAP kinases also regulate intracellular signalingpathways. They mediate signal transduction from the cell surface to thenucleus via phosphorylation cascades. Several subgroups have beenidentified, and each manifests different substrate specificities andresponds to distinct extracellular stimuli (Egan, S. E. and Weinberg, R.A. (1993) Nature 365:781-783). MAP kinase signaling pathways are presentin mammalian cells as well as in yeast. The extracellular stimuli thatactivate mammalian pathways include epidermal growth factor (EGF),ultraviolet light, hyperosmolar medium, heat shock, endotoxiclipopolysaccharide (LPS), and pro-inflammatory cytokines such as tumornecrosis factor (TNF) and interleukin-1(IL-1).

[0010] PRK (proliferation-related kinase) is a serum/cytokine inducibleSTK that is involved in regulation of the cell cycle and cellproliferation in human megakaroytic cells (Li, B. et al. (1996) J. Biol.Chem. 271:19402-8). PRK is related to the polo (derived from humans pologene) family of STKs implicated in cell division. PRK is downregulatedin lung tumor tissue and may be a proto-oncogene whose deregulatedexpression in normal tissue leads to oncogenic transformation. AlteredMAP kinase expression is implicated in a variety of disease conditionsincluding cancer, inflammation, immune disorders, and disordersaffecting growth and development.

[0011] The cyclin-dependent protein kinases (CDKs) are another group ofSTKs that control the progression of cells through the cell cycle.Cyclins are small regulatory proteins that act by binding to andactivating CDKs that then trigger various phases of the cell cycle byphosphorylating and activating selected proteins involved in the mitoticprocess. CDKs are unique in that they require multiple inputs to becomeactivated. In addition to the binding of cyclin, CDK activation requiresthe phosphorylation of a specific threonine residue and thedephosphorylation of a specific tyrosine residue.

[0012] Protein tyrosine kinases, PTKs, specifically phosphorylatetyrosine residues on their target proteins and may be divided intotransmembrane, receptor PTKs and nontransmembrane, non-receptor PTKs.Transmembrane protein-tyrosine kinases are receptors for most growthfactors. Binding of growth factor to the receptor activates the transferof a phosphate group from ATP to selected tyrosine side chains of thereceptor and other specific proteins. Growth factors (GF) associatedwith receptor PTKs include; epidermal GF, platelet-derived GF,fibroblast GF, hepatocyte GF, insulin and insulin-like GFs, nerve GF,vascular endothelial GF, and macrophage colony stimulating factor.

[0013] Non-receptor PTKs lack transmembrane regions and, instead, formcomplexes with the intracellular regions of cell surface receptors. Suchreceptors that function through non-receptor PTKs include those forcytokines, hormones (growth hormone and prolactin) and antigen-specificreceptors on T and B lymphocytes.

[0014] Many of these PTKs were first identified as the products ofmutant oncogenes in cancer cells where their activation was no longersubject to normal cellular controls. In fact, about one third of theknown oncogenes encode PTKs, and it is well known that cellulartransformation (oncogenesis) is often accompanied by increased tyrosinephosphorylation activity (Carbonneau H and Tonks N K (1992) Annu. Rev.Cell. Biol. 8:463-93). Regulation of PTK activity may therefore be animportant strategy in controlling some types of cancer.

[0015] Serine/Threonine Protein Kinases

[0016] The novel human protein, and encoding gene, provided by thepresent invention is related to the serine/threonine protein kinasesubfamily, and shows the highest degree of similarity to striatedmuscle-specific serine/threonine protein kinases.

[0017] At least four isoforms related to striated muscle-specificserine/threonine protein kinases have been identified in the art: a1.4-kb mRNA (aortic preferentially expressed gene (APEG)-1) expressed invascular smooth muscle cells and down-regulated by vascular injury; 9-kbstriated preferentially expressed gene (SPEG)alpha and 11-kb SPEGbeta,both of which are expressed in skeletal muscle and heart; and a 4-kbbrain preferentially expressed gene (BPEG), which is expressed in thebrain and aorta. All four isoforms share the middle three of the fiveexons of APEG-1 but have different alternative spliced 5′- and 3′-ends(Hsieh et al., J. Biol. Chem. 275 (47), 36966-36973 (2000)). SPEGbetacontains two serine/threonine kinase domains and is homologous to myosinlight chain kinase proteins. At least one of the kinase domains inSPEGbeta is active and able to autophosphorylate (Hsieh et al., J. Biol.Chem. 275 (47), 36966-36973 (2000)). Hsieh et al (J. Biol. Chem. 275(47), 36966-36973 (2000)) showed that expression of SPEGalpha andSPEGbeta is developmentally regulated in the striated muscle duringC2C12 myoblast to myotube differentiation in vitro and cardiomyocytematuration in vivo. Hsieh et al suggested that this developmentalregulation indicates that both SPEGalpha and SPEGbeta can serve assensitive markers for striated muscle differentiation and that bothSPEGalpha and SPEGbeta may play important roles in adult striated musclefunction.

[0018] Kinase proteins, particularly members of the serine/threonineprotein kinase subfamily, are a major target for drug action anddevelopment. Accordingly, it is valuable to the field of pharmaceuticaldevelopment to identify and characterize previously unknown members ofthis subfamily of kinase proteins. The present invention advances thestate of the art by providing previously unidentified human kinaseproteins that have homology to members of the serine/threonine proteinkinase subfamily.

SUMMARY OF THE INVENTION

[0019] The present invention is based in part on the identification ofamino acid sequences of human kinase peptides and proteins that arerelated to the serine/threonine protein kinase subfamily, as well asallelic variants and other mammalian orthologs thereof. These uniquepeptide sequences, and nucleic acid sequences that encode thesepeptides, can be used as models for the development of human therapeutictargets, aid in the identification of therapeutic proteins, and serve astargets for the development of human therapeutic agents that modulatekinase activity in cells and tissues that express the kinase.Experimental data as provided in FIG. 1 indicates expression in adultand fetal brain (including astrocytoma and neuroblastoma cells),lung/spleen, and squamous cell carcinoma (skin).

DESCRIPTION OF THE FIGURE SHEETS

[0020]FIG. 1 provides the nucleotide sequence of a transcript sequencethat encodes the kinase protein of the present invention. (SEQ ID NO:1)In addition, structure and functional information is provided, such asATG start, stop and tissue distribution, where available, that allowsone to readily determine specific uses of inventions based on thismolecular sequence. Experimental data as provided in FIG. 1 indicatesexpression in adult and fetal brain (including astrocytoma andneuroblastoma cells), lung/spleen, and squamous cell carcinoma (skin).

[0021]FIG. 2 provides the predicted amino acid sequence of the kinase ofthe present invention. (SEQ ID NO:2) In addition structure andfunctional information such as protein family, function, andmodification sites is provided where available, allowing one to readilydetermine specific uses of inventions based on this molecular sequence.

[0022]FIG. 3 provides genomic sequences that span the gene encoding thekinase protein of the present invention. (SEQ ID NO:3) In additionstructure and functional information, such as intron/exon structure,promoter location, etc., is provided where available, allowing one toreadily determine specific uses of inventions based on this molecularsequence. As illustrated in FIG. 3, SNPs were identified at 39 differentnucleotide positions, including 2 non-synonymous coding SNPs.

DETAILED DESCRIPTION OF THE INVENTION

[0023] General Description The present invention is based on thesequencing of the human genome. During the sequencing and assembly ofthe human genome, analysis of the sequence information revealedpreviously unidentified fragments of the human genome that encodepeptides that share structural and/or sequence homology toprotein/peptide/domains identified and characterized within the art asbeing a kinase protein or part of a kinase protein and are related tothe serine/threonine protein kinase subfamily. Utilizing thesesequences, additional genomic sequences were assembled and transcriptand/or cDNA sequences were isolated and characterized. Based on thisanalysis, the present invention provides amino acid sequences of humankinase peptides and proteins that are related to the serine/threonineprotein kinase subfamily, nucleic acid sequences in the form oftranscript sequences, cDNA sequences and/or genomic sequences thatencode these kinase peptides and proteins, nucleic acid variation(allelic information), tissue distribution of expression, andinformation about the closest art known protein/peptide/domain that hasstructural or sequence homology to the kinase of the present invention.

[0024] In addition to being previously unknown, the peptides that areprovided in the present invention are selected based on their ability tobe used for the development of commercially important products andservices. Specifically, the present peptides are selected based onhomology and/or structural relatedness to known kinase proteins of theserine/threonine protein kinase subfamily and the expression patternobserved. Experimental data as provided in FIG. 1 indicates expressionin adult and fetal brain (including astrocytoma and neuroblastomacells), lung/spleen, and squamous cell carcinoma (skin). The art hasclearly established the commercial importance of members of this familyof proteins and proteins that have expression patterns similar to thatof the present gene. Some of the more specific features of the peptidesof the present invention, and the uses thereof, are described herein,particularly in the Background of the Invention and in the annotationprovided in the Figures, and/or are known within the art for each of theknown serine/threonine protein kinase family or subfamily of kinaseproteins.

[0025] Specific Embodiments

[0026] Peptide Molecules

[0027] The present invention provides nucleic acid sequences that encodeprotein molecules that have been identified as being members of thekinase family of proteins and are related to the serine/threonineprotein kinase subfamily (protein sequences are provided in FIG. 2,transcript/cDNA sequences are provided in FIG. 1 and genomic sequencesare provided in FIG. 3). The peptide sequences provided in FIG. 2, aswell as the obvious variants described herein, particularly allelicvariants as identified herein and using the information in FIG. 3, willbe referred herein as the kinase peptides of the present invention,kinase peptides, or peptides/proteins of the present invention.

[0028] The present invention provides isolated peptide and proteinmolecules that consist of, consist essentially of, or comprise the aminoacid sequences of the kinase peptides disclosed in the FIG. 2, (encodedby the nucleic acid molecule shown in FIG. 1, transcript/cDNA or FIG. 3,genomic sequence), as well as all obvious variants of these peptidesthat are within the art to make and use. Some of these variants aredescribed in detail below.

[0029] As used herein, a peptide is said to be “isolated” or “purified”when it is substantially free of cellular material or free of chemicalprecursors or other chemicals. The peptides of the present invention canbe purified to homogeneity or other degrees of purity. The level ofpurification will be based on the intended use. The critical feature isthat the preparation allows for the desired function of the peptide,even if in the presence of considerable amounts of other components (thefeatures of an isolated nucleic acid molecule is discussed below).

[0030] In some uses, “substantially free of cellular material” includespreparations of the peptide having less than about 30% (by dry weight)other proteins (i.e., contaminating protein), less than about 20% otherproteins, less than about 10% other proteins, or less than about 5%other proteins. When the peptide is recombinantly produced, it can alsobe substantially free of culture medium, i.e., culture medium representsless than about 20% of the volume of the protein preparation.

[0031] The language “substantially free of chemical precursors or otherchemicals” includes preparations of the peptide in which it is separatedfrom chemical precursors or other chemicals that are involved in itssynthesis. In one embodiment, the language “substantially free ofchemical precursors or other chemicals” includes preparations of thekinase peptide having less than about 30% (by dry weight) chemicalprecursors or other chemicals, less than about 20% chemical precursorsor other chemicals, less than about 10% chemical precursors or otherchemicals, or less than about 5% chemical precursors or other chemicals.

[0032] The isolated kinase peptide can be purified from cells thatnaturally express it, purified from cells that have been altered toexpress it (recombinant), or synthesized using known protein synthesismethods. Experimental data as provided in FIG. 1 indicates expression inadult and fetal brain (including astrocytoma and neuroblastoma cells),lung/spleen, and squamous cell carcinoma (skin). For example, a nucleicacid molecule encoding the kinase peptide is cloned into an expressionvector, the expression vector introduced into a host cell and theprotein expressed in the host cell. The protein can then be isolatedfrom the cells by an appropriate purification scheme using standardprotein purification techniques. Many of these techniques are describedin detail below.

[0033] Accordingly, the present invention provides proteins that consistof the amino acid sequences provided in FIG. 2 (SEQ ID NO:2), forexample, proteins encoded by the transcript/cDNA nucleic acid sequencesshown in FIG. 1 (SEQ ID NO:1) and the genomic sequences provided in FIG.3 (SEQ ID NO:3). The amino acid sequence of such a protein is providedin FIG. 2. A protein consists of an amino acid sequence when the aminoacid sequence is the final amino acid sequence of the protein.

[0034] The present invention further provides proteins that consistessentially of the amino acid sequences provided in FIG. 2 (SEQ IDNO:2), for example, proteins encoded by the transcript/cDNA nucleic acidsequences shown in FIG. 1 (SEQ ID NO:1) and the genomic sequencesprovided in FIG. 3 (SEQ ID NO:3). A protein consists essentially of anamino acid sequence when such an amino acid sequence is present withonly a few additional amino acid residues, for example from about 1 toabout 100 or so additional residues, typically from 1 to about 20additional residues in the final protein.

[0035] The present invention further provides proteins that comprise theamino acid sequences provided in FIG. 2 (SEQ ID NO:2), for example,proteins encoded by the transcript/cDNA nucleic acid sequences shown inFIG. 1 (SEQ ID NO:1) and the genomic sequences provided in FIG. 3 (SEQID NO:3). A protein comprises an amino acid sequence when the amino acidsequence is at least part of the final amino acid sequence of theprotein. In such a fashion, the protein can be only the peptide or haveadditional amino acid molecules, such as amino acid residues (contiguousencoded sequence) that are naturally associated with it or heterologousamino acid residues/peptide sequences. Such a protein can have a fewadditional amino acid residues or can comprise several hundred or moreadditional amino acids. The preferred classes of proteins that arecomprised of the kinase peptides of the present invention are thenaturally occurring mature proteins. A brief description of how varioustypes of these proteins can be made/isolated is provided below.

[0036] The kinase peptides of the present invention can be attached toheterologous sequences to form chimeric or fusion proteins. Suchchimeric and fusion proteins comprise a kinase peptide operativelylinked to a heterologous protein having an amino acid sequence notsubstantially homologous to the kinase peptide. “Operatively linked”indicates that the kinase peptide and the heterologous protein are fusedin-frame. The heterologous protein can be fused to the N-terminus orC-terminus of the kinase peptide.

[0037] In some uses, the fusion protein does not affect the activity ofthe kinase peptide per se. For example, the fusion protein can include,but is not limited to, enzymatic fusion proteins, for examplebeta-galactosidase fusions, yeast two-hybrid GAL fusions, poly-Hisfusions, MYC-tagged, HI-tagged and Ig fusions. Such fusion proteins,particularly poly-His fusions, can facilitate the purification ofrecombinant kinase peptide. In certain host cells (e.g., mammalian hostcells), expression and/or secretion of a protein can be increased byusing a heterologous signal sequence.

[0038] A chimeric or fusion protein can be produced by standardrecombinant DNA techniques. For example, DNA fragments coding for thedifferent protein sequences are ligated together in-frame in accordancewith conventional techniques. In another embodiment, the fusion gene canbe synthesized by conventional techniques including automated DNAsynthesizers. Alternatively, PCR amplification of gene fragments can becarried out using anchor primers which give rise to complementaryoverhangs between two consecutive gene fragments which can subsequentlybe annealed and re-amplified to generate a chimeric gene sequence (seeAusubel et al., Current Protocols in Molecular Biology, 1992). Moreover,many expression vectors are commercially available that already encode afusion moiety (e.g., a GST protein). A kinase peptide-encoding nucleicacid can be cloned into such an expression vector such that the fusionmoiety is linked in-frame to the kinase peptide.

[0039] As mentioned above, the present invention also provides andenables obvious variants of the amino acid sequence of the proteins ofthe present invention, such as naturally occurring mature forms of thepeptide, allelic/sequence variants of the peptides, non-naturallyoccurring recombinantly derived variants of the peptides, and orthologsand paralogs of the peptides. Such variants can readily be generatedusing art-known techniques in the fields of recombinant nucleic acidtechnology and protein biochemistry. It is understood, however, thatvariants exclude any amino acid sequences disclosed prior to theinvention.

[0040] Such variants can readily be identified/made using moleculartechniques and the sequence information disclosed herein. Further, suchvariants can readily be distinguished from other peptides based onsequence and/or structural homology to the kinase peptides of thepresent invention. The degree of homology/identity present will be basedprimarily on whether the peptide is a functional variant ornon-functional variant, the amount of divergence present in the paralogfamily and the evolutionary distance between the orthologs.

[0041] To determine the percent identity of two amino acid sequences ortwo nucleic acid sequences, the sequences are aligned for optimalcomparison purposes (e.g., gaps can be introduced in one or both of afirst and a second amino acid or nucleic acid sequence for optimalalignment and non-homologous sequences can be disregarded for comparisonpurposes). In a preferred embodiment, at least 30%, 40%, 50%, 60%, 70%,80%, or 90% or more of the length of a reference sequence is aligned forcomparison purposes. The amino acid residues or nucleotides atcorresponding amino acid positions or nucleotide positions are thencompared. When a position in the first sequence is occupied by the sameamino acid residue or nucleotide as the corresponding position in thesecond sequence, then the molecules are identical at that position (asused herein amino acid or nucleic acid “identity” is equivalent to aminoacid or nucleic acid “homology”). The percent identity between the twosequences is a function of the number of identical positions shared bythe sequences, taking into account the number of gaps, and the length ofeach gap, which need to be introduced for optimal alignment of the twosequences.

[0042] The comparison of sequences and determination of percent identityand similarity between two sequences can be accomplished using amathematical algorithm. (Computational Molecular Biology, Lesk, A. M.,ed., Oxford University Press, New York, 1988; Biocomputing: Informaticsand Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993;Computer Analysis of Sequence Data, Part 1, Griffin, A. M., and Griffin,H. G., eds., Humana Press, New Jersey, 1994; Sequence Analysis inMolecular Biology, von Heinje, G., Academic Press, 1987; and SequenceAnalysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press,New York, 1991). In a preferred embodiment, the percent identity betweentwo amino acid sequences is determined using the Needleman and Wunsch(J. Mol. Biol. (48):444-453 (1970)) algorithm which has beenincorporated into the GAP program in the GCG software package (availableat http://www.gcg.com), using either a Blossom 62 matrix or a PAM 250matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a lengthweight of 1, 2, 3, 4, 5, or 6. In yet another preferred embodiment, thepercent identity between two nucleotide sequences is determined usingthe GAP program in the GCG software package (Devereux, J., et al.,Nucleic Acids Res. 12(1):387 (1984)) (available at http://www.gcg.com),using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80and a length weight of 1, 2, 3, 4, 5, or 6. In another embodiment, thepercent identity between two amino acid or nucleotide sequences isdetermined using the algorithm of E. Myers and W. Miller (CABIOS,4:11-17 (1989)) which has been incorporated into the ALIGN program(version 2.0), using a PAM 120 weight residue table, a gap lengthpenalty of 12 and a gap penalty of 4.

[0043] The nucleic acid and protein sequences of the present inventioncan further be used as a “query sequence” to perform a search againstsequence databases to, for example, identify other family members orrelated sequences. Such searches can be performed using the NBLAST andXBLAST programs (version 2.0) of Altschul, et al. (J. Mol. Biol.215:403-10 (1990)). BLAST nucleotide searches can be performed with theNBLAST program, score=100, wordlength=12 to obtain nucleotide sequenceshomologous to the nucleic acid molecules of the invention. BLAST proteinsearches can be performed with the XBLAST program, score=50,wordlength=3 to obtain amino acid sequences homologous to the proteinsof the invention. To obtain gapped alignments for comparison purposes,Gapped BLAST can be utilized as described in Altschul et al. (NucleicAcids Res. 25(17):3389-3402 (1997)). When utilizing BLAST and gappedBLAST programs, the default parameters of the respective programs (e.g.,XBLAST and NBLAST) can be used.

[0044] Full-length pre-processed forms, as well as mature processedforms, of proteins that comprise one of the peptides of the presentinvention can readily be identified as having complete sequence identityto one of the kinase peptides of the present invention as well as beingencoded by the same genetic locus as the kinase peptide provided herein.As indicated in FIG. 3, the map position was determined to be on humanchromosome 2.

[0045] Allelic variants of a kinase peptide can readily be identified asbeing a human protein having a high degree (significant) of sequencehomology/identity to at least a portion of the kinase peptide as well asbeing encoded by the same genetic locus as the kinase peptide providedherein. Genetic locus can readily be determined based on the genomicinformation provided in FIG. 3, such as the genomic sequence mapped tothe reference human. As indicated in FIG. 3, the map position wasdetermined to be on human chromosome 2. As used herein, two proteins (ora region of the proteins) have significant homology when the amino acidsequences are typically at least about 70-80%, 80-90%, and moretypically at least about 90-95% or more homologous. A significantlyhomologous amino acid sequence, according to the present invention, willbe encoded by a nucleic acid sequence that will hybridize to a kinasepeptide encoding nucleic acid molecule under stringent conditions asmore fully described below.

[0046]FIG. 3 provides information on SNPs that have been found in thegene encoding the kinase proteins of the present invention. SNPs wereidentified at 39 different nucleotide positions, including five SNPs incoding regions, two of which (at nucleotide positions 52048 and 58826)change the encoded amino acid. The changes in the amino acid sequencethat these SNPs cause is indicated in FIG. 3 and can readily bedetermined using the universal genetic code and the protein sequenceprovided in FIG. 2 as a reference.

[0047] Paralogs of a kinase peptide can readily be identified as havingsome degree of significant sequence homology/identity to at least aportion of the kinase peptide, as being encoded by a gene from humans,and as having similar activity or function. Two proteins will typicallybe considered paralogs when the amino acid sequences are typically atleast about 60% or greater, and more typically at least about 70% orgreater homology through a given region or domain. Such paralogs will beencoded by a nucleic acid sequence that will hybridize to a kinasepeptide encoding nucleic acid molecule under moderate to stringentconditions as more fully described below.

[0048] Orthologs of a kinase peptide can readily be identified as havingsome degree of significant sequence homology/identity to at least aportion of the kinase peptide as well as being encoded by a gene fromanother organism. Preferred orthologs will be isolated from mammals,preferably primates, for the development of human therapeutic targetsand agents. Such orthologs will be encoded by a nucleic acid sequencethat will hybridize to a kinase peptide encoding nucleic acid moleculeunder moderate to stringent conditions, as more fully described below,depending on the degree of relatedness of the two organisms yielding theproteins.

[0049] Non-naturally occurring variants of the kinase peptides of thepresent invention can readily be generated using recombinant techniques.Such variants include, but are not limited to deletions, additions andsubstitutions in the amino acid sequence of the kinase peptide. Forexample, one class of substitutions are conserved amino acidsubstitution. Such substitutions are those that substitute a given aminoacid in a kinase peptide by another amino acid of like characteristics.Typically seen as conservative substitutions are the replacements, onefor another, among the aliphatic amino acids Ala, Val, Leu, and Ile;interchange of the hydroxyl residues Ser and Thr; exchange of the acidicresidues Asp and Glu; substitution between the amide residues Asn andGln; exchange of the basic residues Lys and Arg; and replacements amongthe aromatic residues Phe and Tyr. Guidance concerning which amino acidchanges are likely to be phenotypically silent are found in Bowie etal., Science 247:1306-1310 (1990).

[0050] Variant kinase peptides can be filly functional or can lackfunction in one or more activities, e.g. ability to bind substrate,ability to phosphorylate substrate, ability to mediate signaling, etc.Fully functional variants typically contain only conservative variationor variation in non-critical residues or in non-critical regions. FIG. 2provides the result of protein analysis and can be used to identifycritical domains/regions. Functional variants can also containsubstitution of similar amino acids that result in no change or aninsignificant change in function. Alternatively, such substitutions maypositively or negatively affect function to some degree.

[0051] Non-functional variants typically contain one or morenon-conservative amino acid substitutions, deletions, insertions,inversions, or truncation or a substitution, insertion, inversion, ordeletion in a critical residue or critical region.

[0052] Amino acids that are essential for function can be identified bymethods known in the art, such as site-directed mutagenesis oralanine-scanning mutagenesis (Cunningham et al., Science 244:1081-1085(1989)), particularly using the results provided in FIG. 2. The latterprocedure introduces single alanine mutations at every residue in themolecule. The resulting mutant molecules are then tested for biologicalactivity such as kinase activity or in assays such as an in vitroproliferative activity. Sites that are critical for bindingpartner/substrate binding can also be determined by structural analysissuch as crystallization, nuclear magnetic resonance or photoaffinitylabeling (Smith et al., J. Mol. Biol. 224:899-904 (1992); de Vos et al.Science 255:306-312 (1992)).

[0053] The present invention further provides fragments of the kinasepeptides, in addition to proteins and peptides that comprise and consistof such fragments, particularly those comprising the residues identifiedin FIG. 2. The fragments to which the invention pertains, however, arenot to be construed as encompassing fragments that may be disclosedpublicly prior to the present invention.

[0054] As used herein, a fragment comprises at least 8, 10, 12, 14, 16,or more contiguous amino acid residues from a kinase peptide. Suchfragments can be chosen based on the ability to retain one or more ofthe biological activities of the kinase peptide or could be chosen forthe ability to perform a function, e.g. bind a substrate or act as animmunogen. Particularly important fragments are biologically activefragments, peptides that are, for example, about 8 or more amino acidsin length. Such fragments will typically comprise a domain or motif ofthe kinase peptide, e.g., active site, a transmembrane domain or asubstrate-binding domain. Further, possible fragments include, but arenot limited to, domain or motif containing fragments, soluble peptidefragments, and fragments containing immunogenic structures. Predicteddomains and functional sites are readily identifiable by computerprograms well known and readily available to those of skill in the art(e.g., PROSITE analysis). The results of one such analysis are providedin FIG. 2.

[0055] Polypeptides often contain amino acids other than the 20 aminoacids commonly referred to as the 20 naturally occurring amino acids.Further, many amino acids, including the terminal amino acids, may bemodified by natural processes, such as processing and otherpost-translational modifications, or by chemical modification techniqueswell known in the art. Common modifications that occur naturally inkinase peptides are described in basic texts, detailed monographs, andthe research literature, and they are well known to those of skill inthe art (some of these features are identified in FIG. 2).

[0056] Known modifications include, but are not limited to, acetylation,acylation, ADP-ribosylation, amidation, covalent attachment of flavin,covalent attachment of a heme moiety, covalent attachment of anucleotide or nucleotide derivative, covalent attachment of a lipid orlipid derivative, covalent attachment of phosphotidylinositol,cross-linking, cyclization, disulfide bond formation, demethylation,formation of covalent crosslinks, formation of cystine, formation ofpyroglutamate, formylation, gamma carboxylation, glycosylation, GPIanchor formation, hydroxylation, iodination, methylation,myristoylation, oxidation, proteolytic processing, phosphorylation,prenylation, racemization, selenoylation, sulfation, transfer-RNAmediated addition of amino acids to proteins such as arginylation, andubiquitination.

[0057] Such modifications are well known to those of skill in the artand have been described in great detail in the scientific literature.Several particularly common modifications, glycosylation, lipidattachment, sulfation, gamma-carboxylation of glutamic acid residues,hydroxylation and ADP-ribosylation, for instance, are described in mostbasic texts, such as Proteins—Structure and Molecular Properties, 2ndEd., T. E. Creighton, W. H. Freeman and Company, New York (1993). Manydetailed reviews are available on this subject, such as by Wold, F.,Posttranslational Covalent Modification of Proteins, B. C. Johnson, Ed.,Academic Press, New York 1-12 (1983); Seifter et al. (Meth. Enzymol.182: 626-646 (1990)) and Rattan et al. (Ann. N.Y Acad. Sci. 663:48-62(1992)).

[0058] Accordingly, the kinase peptides of the present invention alsoencompass derivatives or analogs in which a substituted amino acidresidue is not one encoded by the genetic code, in which a substituentgroup is included, in which the mature kinase peptide is fused withanother compound, such as a compound to increase the half-life of thekinase peptide (for example, polyethylene glycol), or in which theadditional amino acids are fused to the mature kinase peptide, such as aleader or secretory sequence or a sequence for purification of themature kinase peptide or a pro-protein sequence.

[0059] Protein/Peptide Uses

[0060] The proteins of the present invention can be used in substantialand specific assays related to the functional information provided inthe Figures; to raise antibodies or to elicit another immune response;as a reagent (including the labeled reagent) in assays designed toquantitatively determine levels of the protein (or its binding partneror ligand) in biological fluids; and as markers for tissues in which thecorresponding protein is preferentially expressed (either constitutivelyor at a particular stage of tissue differentiation or development or ina disease state). Where the protein binds or potentially binds toanother protein or ligand (such as, for example, in a kinase-effectorprotein interaction or kinase-ligand interaction), the protein can beused to identify the binding partner/ligand so as to develop a system toidentify inhibitors of the binding interaction. Any or all of these usesare capable of being developed into reagent grade or kit format forcommercialization as commercial products.

[0061] Methods for performing the uses listed above are well known tothose skilled in the art. References disclosing such methods include“Molecular Cloning: A Laboratory Manual”, 2d ed., Cold Spring HarborLaboratory Press, Sambrook, J., E. F. Fritsch and T. Maniatis eds.,1989, and “Methods in Enzymology: Guide to Molecular CloningTechniques”, Academic Press, Berger, S. L. and A. R. Kimmel eds., 1987.

[0062] The potential uses of the peptides of the present invention arebased primarily on the source of the protein as well as the class/actionof the protein. For example, kinases isolated from humans and theirhuman/mammalian orthologs serve as targets for identifying agents foruse in mammalian therapeutic applications, e.g. a human drug,particularly in modulating a biological or pathological response in acell or tissue that expresses the kinase. Experimental data as providedin FIG. 1 indicates that kinase proteins of the present invention areexpressed in adult and fetal brain (including astrocytoma andneuroblastoma cells), lung/spleen, and squamous cell carcinoma (skin),as indicated by virtual northern blot analysis. A large percentage ofpharmaceutical agents are being developed that modulate the activity ofkinase proteins, particularly members of the serine/threonine proteinkinase subfamily (see Background of the Invention). The structural andfunctional information provided in the Background and Figures providespecific and substantial uses for the molecules of the presentinvention, particularly in combination with the expression informationprovided in FIG. 1. Experimental data as provided in FIG. 1 indicatesexpression in adult and fetal brain (including astrocytoma andneuroblastoma cells), lung/spleen, and squamous cell carcinoma (skin).Such uses can readily be determined using the information providedherein, that which is known in the art, and routine experimentation.

[0063] The proteins of the present invention (including variants andfragments that may have been disclosed prior to the present invention)are useful for biological assays related to kinases that are related tomembers of the serine/threonine protein kinase subfamily. Such assaysinvolve any of the known kinase functions or activities or propertiesuseful for diagnosis and treatment of kinase-related conditions that arespecific for the subfamily of kinases that the one of the presentinvention belongs to, particularly in cells and tissues that express thekinase. Experimental data as provided in FIG. 1 indicates that kinaseproteins of the present invention are expressed in adult and fetal brain(including astrocytoma and neuroblastoma cells), lung/spleen, andsquamous cell carcinoma (skin), as indicated by virtual northern blotanalysis.

[0064] The proteins of the present invention are also useful in drugscreening assays, in cell-based or cell-free systems. Cell-based systemscan be native, i.e., cells that normally express the kinase, as a biopsyor expanded in cell culture. Experimental data as provided in FIG. 1indicates expression in adult and fetal brain (including astrocytoma andneuroblastoma cells), lung/spleen, and squamous cell carcinoma (skin).In an alternate embodiment, cell-based assays involve recombinant hostcells expressing the kinase protein.

[0065] The polypeptides can be used to identify compounds that modulatekinase activity of the protein in its natural state or an altered formthat causes a specific disease or pathology associated with the kinase.Both the kinases of the present invention and appropriate variants andfragments can be used in high-throughput screens to assay candidatecompounds for the ability to bind to the kinase. These compounds can befurther screened against a functional kinase to determine the effect ofthe compound on the kinase activity. Further, these compounds can betested in animal or invertebrate systems to determineactivity/effectiveness. Compounds can be identified that activate(agonist) or inactivate (antagonist) the kinase to a desired degree.

[0066] Further, the proteins of the present invention can be used toscreen a compound for the ability to stimulate or inhibit interactionbetween the kinase protein and a molecule that normally interacts withthe kinase protein, e.g. a substrate or a component of the signalpathway that the kinase protein normally interacts (for example, anotherkinase). Such assays typically include the steps of combining the kinaseprotein with a candidate compound under conditions that allow the kinaseprotein, or fragment, to interact with the target molecule, and todetect the formation of a complex between the protein and the target orto detect the biochemical consequence of the interaction with the kinaseprotein and the target, such as any of the associated effects of signaltransduction such as protein phosphorylation, cAMP turnover, andadenylate cyclase activation, etc.

[0067] Candidate compounds include, for example, 1) peptides such assoluble peptides, including Ig-tailed fusion peptides and members ofrandom peptide libraries (see, e.g., Lam et al., Nature 354:82-84(1991); Houghten et al, Nature 354:84-86 (1991)) and combinatorialchemistry-derived molecular libraries made of D- and/or L-configurationamino acids; 2) phosphopeptides (e.g., members of random and partiallydegenerate, directed phosphopeptide libraries, see, e.g., Songyang etal., Cell 72:767-778 (1993)); 3) antibodies (e.g., polyclonal,monoclonal, humanized, anti-idiotypic, chimeric, and single chainantibodies as well as Fab, F(ab′)₂, Fab expression library fragments,and epitope-binding fragments of antibodies); and 4) small organic andinorganic molecules (e.g., molecules obtained from combinatorial andnatural product libraries).

[0068] One candidate compound is a soluble fragment of the receptor thatcompetes for substrate binding. Other candidate compounds include mutantkinases or appropriate fragments containing mutations that affect kinasefunction and thus compete for substrate. Accordingly, a fragment thatcompetes for substrate, for example with a higher affinity, or afragment that binds substrate but does not allow release, is encompassedby the invention.

[0069] The invention further includes other end point assays to identifycompounds that modulate (stimulate or inhibit) kinase activity. Theassays typically involve an assay of events in the signal transductionpathway that indicate kinase activity. Thus, the phosphorylation of asubstrate, activation of a protein, a change in the expression of genesthat are up- or down-regulated in response to the kinase proteindependent signal cascade can be assayed.

[0070] Any of the biological or biochemical functions mediated by thekinase can be used as an endpoint assay. These include all of thebiochemical or biochemical/biological events described herein, in thereferences cited herein, incorporated by reference for these endpointassay targets, and other functions known to those of ordinary skill inthe art or that can be readily identified using the information providedin the Figures, particularly FIG. 2. Specifically, a biological functionof a cell or tissues that expresses the kinase can be assayed.Experimental data as provided in FIG. 1 indicates that kinase proteinsof the present invention are expressed in adult and fetal brain(including astrocytoma and neuroblastoma cells), lung/spleen, andsquamous cell carcinoma (skin), as indicated by virtual northern blotanalysis.

[0071] Binding and/or activating compounds can also be screened by usingchimeric kinase proteins in which the amino terminal extracellulardomain, or parts thereof, the entire transmembrane domain or subregions,such as any of the seven transmembrane segments or any of theintracellular or extracellular loops and the carboxy terminalintracellular domain, or parts thereof, can be replaced by heterologousdomains or subregions. For example, a substrate-binding region can beused that interacts with a different substrate then that which isrecognized by the native kinase. Accordingly, a different set of signaltransduction components is available as an end-point assay foractivation. This allows for assays to be performed in other than thespecific host cell from which the kinase is derived.

[0072] The proteins of the present invention are also useful incompetition binding assays in methods designed to discover compoundsthat interact with the kinase (e.g. binding partners and/or ligands).Thus, a compound is exposed to a kinase polypeptide under conditionsthat allow the compound to bind or to otherwise interact with thepolypeptide. Soluble kinase polypeptide is also added to the mixture. Ifthe test compound interacts with the soluble kinase polypeptide, itdecreases the amount of complex formed or activity from the kinasetarget. This type of assay is particularly useful in cases in whichcompounds are sought that interact with specific regions of the kinase.Thus, the soluble polypeptide that competes with the target kinaseregion is designed to contain peptide sequences corresponding to theregion of interest.

[0073] To perform cell free drug screening assays, it is sometimesdesirable to immobilize either the kinase protein, or fragment, or itstarget molecule to facilitate separation of complexes from uncomplexedforms of one or both of the proteins, as well as to accommodateautomation of the assay.

[0074] Techniques for immobilizing proteins on matrices can be used inthe drug screening assays.

[0075] In one embodiment, a fusion protein can be provided which adds adomain that allows the protein to be bound to a matrix. For example,glutathione-S-transferase fusion proteins can be adsorbed ontoglutathione sepharose beads (Sigma Chemical, St. Louis, Mo.) orglutathione derivatized microtitre plates, which are then combined withthe cell lysates (e.g., ³⁵S-labeled) and the candidate compound, and themixture incubated under conditions conducive to complex formation (e.g.,at physiological conditions for salt and pH). Following incubation, thebeads are washed to remove any unbound label, and the matrix immobilizedand radiolabel determined directly, or in the supernatant after thecomplexes are dissociated. Alternatively, the complexes can bedissociated from the matrix, separated by SDS-PAGE, and the level ofkinase-binding protein found in the bead fraction quantitated from thegel using standard electrophoretic techniques. For example, either thepolypeptide or its target molecule can be immobilized utilizingconjugation of biotin and streptavidin using techniques well known inthe art. Alternatively, antibodies reactive with the protein but whichdo not interfere with binding of the protein to its target molecule canbe derivatized to the wells of the plate, and the protein trapped in thewells by antibody conjugation. Preparations of a kinase-binding proteinand a candidate compound are incubated in the kinase protein-presentingwells and the amount of complex trapped in the well can be quantitated.Methods for detecting such complexes, in addition to those describedabove for the GST-immobilized complexes, include immunodetection ofcomplexes using antibodies reactive with the kinase protein targetmolecule, or which are reactive with kinase protein and compete with thetarget molecule, as well as enzyme-linked assays which rely on detectingan enzymatic activity associated with the target molecule.

[0076] Agents that modulate one of the kinases of the present inventioncan be identified using one or more of the above assays, alone or incombination. It is generally preferable to use a cell-based or cell freesystem first and then confirm activity in an animal or other modelsystem. Such model systems are well known in the art and can readily beemployed in this context.

[0077] Modulators of kinase protein activity identified according tothese drug screening assays can be used to treat a subject with adisorder mediated by the kinase pathway, by treating cells or tissuesthat express the kinase. Experimental data as provided in FIG. 1indicates expression in adult and fetal brain (including astrocytoma andneuroblastoma cells), lung/spleen, and squamous cell carcinoma (skin).These methods of treatment include the steps of administering amodulator of kinase activity in a pharmaceutical composition to asubject in need of such treatment, the modulator being identified asdescribed herein.

[0078] In yet another aspect of the invention, the kinase proteins canbe used as “bait proteins” in a two-hybrid assay or three-hybrid assay(see, e.g., U.S. Pat. No. 5,283,317; Zervos et al. (1993) Cell72:223-232; Madura et al. (1993) J. Biol. Chem. 268:12046-12054; Bartelet al. (1993) Biotechniques 14:920-924; Iwabuchi et al. (1993) Oncogene8:1693-1696; and Brent WO94/10300), to identify other proteins, whichbind to or interact with the kinase and are involved in kinase activity.Such kinase-binding proteins are also likely to be involved in thepropagation of signals by the kinase proteins or kinase targets as, forexample, downstream elements of a kinase-mediated signaling pathway.Alternatively, such kinase-binding proteins are likely to be kinaseinhibitors.

[0079] The two-hybrid system is based on the modular nature of mosttranscription factors, which consist of separable DNA-binding andactivation domains. Briefly, the assay utilizes two different DNAconstructs. In one construct, the gene that codes for a kinase proteinis fused to a gene encoding the DNA binding domain of a knowntranscription factor (e.g., GAL-4). In the other construct, a DNAsequence, from a library of DNA sequences, that encodes an unidentifiedprotein (“prey” or “sample”) is fused to a gene that codes for theactivation domain of the known transcription factor. If the “bait” andthe “prey” proteins are able to interact, in vivo, forming akinase-dependent complex, the DNA-binding and activation domains of thetranscription factor are brought into close proximity. This proximityallows transcription of a reporter gene (e.g., LacZ) which is operablylinked to a transcriptional regulatory site responsive to thetranscription factor. Expression of the reporter gene can be detectedand cell colonies containing the functional transcription factor can beisolated and used to obtain the cloned gene which encodes the proteinwhich interacts with the kinase protein.

[0080] This invention further pertains to novel agents identified by theabove-described screening assays. Accordingly, it is within the scope ofthis invention to further use an agent identified as described herein inan appropriate animal model. For example, an agent identified asdescribed herein (e.g., a kinase-modulating agent, an antisense kinasenucleic acid molecule, a kinase-specific antibody, or a kinase-bindingpartner) can be used in an animal or other model to determine theefficacy, toxicity, or side effects of treatment with such an agent.Alternatively, an agent identified as described herein can be used in ananimal or other model to determine the mechanism of action of such anagent. Furthermore, this invention pertains to uses of novel agentsidentified by the above-described screening assays for treatments asdescribed herein.

[0081] The kinase proteins of the present invention are also useful toprovide a target for diagnosing a disease or predisposition to diseasemediated by the peptide. Accordingly, the invention provides methods fordetecting the presence, or levels of, the protein (or encoding mRNA) ina cell, tissue, or organism. Experimental data as provided in FIG. 1indicates expression in adult and fetal brain (including astrocytoma andneuroblastoma cells), lung/spleen, and squamous cell carcinoma (skin).The method involves contacting a biological sample with a compoundcapable of interacting with the kinase protein such that the interactioncan be detected. Such an assay can be provided in a single detectionformat or a multi-detection format such as an antibody chip array.

[0082] One agent for detecting a protein in a sample is an antibodycapable of selectively binding to protein. A biological sample includestissues, cells and biological fluids isolated from a subject, as well astissues, cells and fluids present within a subject.

[0083] The peptides of the present invention also provide targets fordiagnosing active protein activity, disease, or predisposition todisease, in a patient having a variant peptide, particularly activitiesand conditions that are known for other members of the family ofproteins to which the present one belongs. Thus, the peptide can beisolated from a biological sample and assayed for the presence of agenetic mutation that results in aberrant peptide. This includes aminoacid substitution, deletion, insertion, rearrangement, (as the result ofaberrant splicing events), and inappropriate post-translationalmodification. Analytic methods include altered electrophoretic mobility,altered tryptic peptide digest, altered kinase activity in cell-based orcell-free assay, alteration in substrate or antibody-binding pattern,altered isoelectric point, direct amino acid sequencing, and any otherof the known assay techniques useful for detecting mutations in aprotein. Such an assay can be provided in a single detection format or amulti-detection format such as an antibody chip array.

[0084] In vitro techniques for detection of peptide include enzymelinked immunosorbent assays (ELISAs), Western blots,immunoprecipitations and immunofluorescence using a detection reagent,such as an antibody or protein binding agent. Alternatively, the peptidecan be detected in vivo in a subject by introducing into the subject alabeled anti-peptide antibody or other types of detection agent. Forexample, the antibody can be labeled with a radioactive marker whosepresence and location in a subject can be detected by standard imagingtechniques. Particularly useful are methods that detect the allelicvariant of a peptide expressed in a subject and methods which detectfragments of a peptide in a sample.

[0085] The peptides are also useful in pharmacogenomic analysis.Pharmacogenomics deal with clinically significant hereditary variationsin the response to drugs due to altered drug disposition and abnormalaction in affected persons. See, e.g., Eichelbaum, M. (Clin. Exp.Pharmacol. Physiol. 23(10-11):983-985(1996)), and Linder, M. W. (Clin.Chem. 43(2):254-266 (1997)). The clinical outcomes of these variationsresult in severe toxicity of therapeutic drugs in certain individuals ortherapeutic failure of drugs in certain individuals as a result ofindividual variation in metabolism. Thus, the genotype of the individualcan determine the way a therapeutic compound acts on the body or the waythe body metabolizes the compound. Further, the activity of drugmetabolizing enzymes effects both the intensity and duration of drugaction. Thus, the pharmacogenomics of the individual permit theselection of effective compounds and effective dosages of such compoundsfor prophylactic or therapeutic treatment based on the individual'sgenotype. The discovery of genetic polymorphisms in some drugmetabolizing enzymes has explained why some patients do not obtain theexpected drug effects, show an exaggerated drug effect, or experienceserious toxicity from standard drug dosages. Polymorphisms can beexpressed in the phenotype of the extensive metabolizer and thephenotype of the poor metabolizer. Accordingly, genetic polymorphism maylead to allelic protein variants of the kinase protein in which one ormore of the kinase functions in one population is different from thosein another population. The peptides thus allow a target to ascertain agenetic predisposition that can affect treatment modality. Thus, in aligand-based treatment, polymorphism may give rise to amino terminalextracellular domains and/or other substrate-binding regions that aremore or less active in substrate binding, and kinase activation.Accordingly, substrate dosage would necessarily be modified to maximizethe therapeutic effect within a given population containing apolymorphism. As an alternative to genotyping, specific polymorphicpeptides could be identified.

[0086] The peptides are also useful for treating a disordercharacterized by an absence of, inappropriate, or unwanted expression ofthe protein. Experimental data as provided in FIG. 1 indicatesexpression in adult and fetal brain (including astrocytoma andneuroblastoma cells), lung/spleen, and squamous cell carcinoma (skin).Accordingly, methods for treatment include the use of the kinase proteinor fragments.

[0087] Antibodies

[0088] The invention also provides antibodies that selectively bind toone of the peptides of the present invention, a protein comprising sucha peptide, as well as variants and fragments thereof. As used herein, anantibody selectively binds a target peptide when it binds the targetpeptide and does not significantly bind to unrelated proteins. Anantibody is still considered to selectively bind a peptide even if italso binds to other proteins that are not substantially homologous withthe target peptide so long as such proteins share homology with afragment or domain of the peptide target of the antibody. In this case,it would be understood that antibody binding to the peptide is stillselective despite some degree of cross-reactivity.

[0089] As used herein, an antibody is defined in terms consistent withthat recognized within the art: they are multi-subunit proteins producedby a mammalian organism in response to an antigen challenge. Theantibodies of the present invention include polyclonal antibodies andmonoclonal antibodies, as well as fragments of such antibodies,including, but not limited to, Fab or F(ab′)₂, and Fv fragments.

[0090] Many methods are known for generating and/or identifyingantibodies to a given target peptide. Several such methods are describedby Harlow, Antibodies, Cold Spring Harbor Press, (1989).

[0091] In general, to generate antibodies, an isolated peptide is usedas an immunogen and is administered to a mammalian organism, such as arat, rabbit or mouse. The full-length protein, an antigenic peptidefragment or a fusion protein can be used. Particularly importantfragments are those covering functional domains, such as the domainsidentified in FIG. 2, and domain of sequence homology or divergenceamongst the family, such as those that can readily be identified usingprotein alignment methods and as presented in the Figures.

[0092] Antibodies are preferably prepared from regions or discretefragments of the kinase proteins. Antibodies can be prepared from anyregion of the peptide as described herein. However, preferred regionswill include those involved in function/activity and/or kinase/bindingpartner interaction. FIG. 2 can be used to identify particularlyimportant regions while sequence alignment can be used to identifyconserved and unique sequence fragments.

[0093] An antigenic fragment will typically comprise at least 8contiguous amino acid residues. The antigenic peptide can comprise,however, at least 10, 12, 14, 16 or more amino acid residues. Suchfragments can be selected on a physical property, such as fragmentscorrespond to regions that are located on the surface of the protein,e.g., hydrophilic regions or can be selected based on sequenceuniqueness (see FIG. 2).

[0094] Detection on an antibody of the present invention can befacilitated by coupling (i.e., physically linking) the antibody to adetectable substance. Examples of detectable substances include variousenzymes, prosthetic groups, fluorescent materials, luminescentmaterials, bioluminescent materials, and radioactive materials. Examplesof suitable enzymes include horseradish peroxidase, alkalinephosphatase, β-galactosidase, or acetylcholinesterase; examples ofsuitable prosthetic group complexes include streptavidin/biotin andavidin/biotin; examples of suitable fluorescent materials includeumbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine,dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; anexample of a luminescent material includes luminol; examples ofbioluminescent materials include luciferase, luciferin, and aequorin,and examples of suitable radioactive material include ¹²⁵I, ¹³¹I, ³⁵S or³H.

[0095] Antibody Uses

[0096] The antibodies can be used to isolate one of the proteins of thepresent invention by standard techniques, such as affinitychromatography or immunoprecipitation. The antibodies can facilitate thepurification of the natural protein from cells and recombinantlyproduced protein expressed in host cells. In addition, such antibodiesare useful to detect the presence of one of the proteins of the presentinvention in cells or tissues to determine the pattern of expression ofthe protein among various tissues in an organism and over the course ofnormal development. Experimental data as provided in FIG. 1 indicatesthat kinase proteins of the present invention are expressed in adult andfetal brain (including astrocytoma and neuroblastoma cells),lung/spleen, and squamous cell carcinoma (skin), as indicated by virtualnorthern blot analysis. Further, such antibodies can be used to detectprotein in situ, in vitro, or in a cell lysate or supernatant in orderto evaluate the abundance and pattern of expression. Also, suchantibodies can be used to assess abnormal tissue distribution orabnormal expression during development or progression of a biologicalcondition. Antibody detection of circulating fragments of the fulllength protein can be used to identify turnover.

[0097] Further, the antibodies can be used to assess expression indisease states such as in active stages of the disease or in anindividual with a predisposition toward disease related to the protein'sfunction. When a disorder is caused by an inappropriate tissuedistribution, developmental expression, level of expression of theprotein, or expressed/processed form, the antibody can be preparedagainst the normal protein. Experimental data as provided in FIG. 1indicates expression in adult and fetal brain (including astrocytoma andneuroblastoma cells), lung/spleen, and squamous cell carcinoma (skin).If a disorder is characterized by a specific mutation in the protein,antibodies specific for this mutant protein can be used to assay for thepresence of the specific mutant protein.

[0098] The antibodies can also be used to assess normal and aberrantsubcellular localization of cells in the various tissues in an organism.Experimental data as provided in FIG. 1 indicates expression in adultand fetal brain (including astrocytoma and neuroblastoma cells),lung/spleen, and squamous cell carcinoma (skin). The diagnostic uses canbe applied, not only in genetic testing, but also in monitoring atreatment modality. Accordingly, where treatment is ultimately aimed atcorrecting expression level or the presence of aberrant sequence andaberrant tissue distribution or developmental expression, antibodiesdirected against the protein or relevant fragments can be used tomonitor therapeutic efficacy.

[0099] Additionally, antibodies are useful in pharmacogenomic analysis.Thus, antibodies prepared against polymorphic proteins can be used toidentify individuals that require modified treatment modalities. Theantibodies are also useful as diagnostic tools as an immunologicalmarker for aberrant protein analyzed by electrophoretic mobility,isoelectric point, tryptic peptide digest, and other physical assaysknown to those in the art.

[0100] The antibodies are also useful for tissue typing. Experimentaldata as provided in FIG. 1 indicates expression in adult and fetal brain(including astrocytoma and neuroblastoma cells), lung/spleen, andsquamous cell carcinoma (skin). Thus, where a specific protein has beencorrelated with expression in a specific tissue, antibodies that arespecific for this protein can be used to identify a tissue type.

[0101] The antibodies are also useful for inhibiting protein function,for example, blocking the binding of the kinase peptide to a bindingpartner such as a substrate. These uses can also be applied in atherapeutic context in which treatment involves inhibiting the protein'sfunction. An antibody can be used, for example, to block binding, thusmodulating (agonizing or antagonizing) the peptides activity. Antibodiescan be prepared against specific fragments containing sites required forfunction or against intact protein that is associated with a cell orcell membrane. See FIG. 2 for structural information relating to theproteins of the present invention.

[0102] The invention also encompasses kits for using antibodies todetect the presence of a protein in a biological sample. The kit cancomprise antibodies such as a labeled or labelable antibody and acompound or agent for detecting protein in a biological sample; meansfor determining the amount of protein in the sample; means for comparingthe amount of protein in the sample with a standard; and instructionsfor use. Such a kit can be supplied to detect a single protein orepitope or can be configured to detect one of a multitude of epitopes,such as in an antibody detection array. Arrays are described in detailbelow for nucleic acid arrays and similar methods have been developedfor antibody arrays.

[0103] Nucleic Acid Molecules

[0104] The present invention further provides isolated nucleic acidmolecules that encode a kinase peptide or protein of the presentinvention (cDNA, transcript and genomic sequence). Such nucleic acidmolecules will consist of, consist essentially of, or comprise anucleotide sequence that encodes one of the kinase peptides of thepresent invention, an allelic variant thereof, or an ortholog or paralogthereof

[0105] As used herein, an “isolated” nucleic acid molecule is one thatis separated from other nucleic acid present in the natural source ofthe nucleic acid. Preferably, an “isolated”. nucleic acid is free ofsequences which naturally flank the nucleic acid (i.e., sequenceslocated at the 5′ and 3′ ends of the nucleic acid) in the genomic DNA ofthe organism from which the nucleic acid is derived. However, there canbe some flanking nucleotide sequences, for example up to about 5KB, 4KB,3KB, 2KB, or 1KB or less, particularly contiguous peptide encodingsequences and peptide encoding sequences within the same gene butseparated by introns in the genomic sequence. The important point isthat the nucleic acid is isolated from remote and unimportant flankingsequences such that it can be subjected to the specific manipulationsdescribed herein such as recombinant expression, preparation of probesand primers, and other uses specific to the nucleic acid sequences.

[0106] Moreover, an “isolated” nucleic acid molecule, such as atranscript/cDNA molecule, can be substantially free of other cellularmaterial, or culture medium when produced by recombinant techniques, orchemical precursors or other chemicals when chemically synthesized.However, the nucleic acid molecule can be fused to other coding orregulatory sequences and still be considered isolated.

[0107] For example, recombinant DNA molecules contained in a vector areconsidered isolated. Further examples of isolated DNA molecules includerecombinant DNA molecules maintained in heterologous host cells orpurified (partially or substantially) DNA molecules in solution.Isolated RNA molecules include in vivo or in vitro. RNA transcripts ofthe isolated DNA molecules of the present invention. Isolated nucleicacid molecules according to the present invention further include suchmolecules produced synthetically.

[0108] Accordingly, the present invention provides nucleic acidmolecules that consist of the nucleotide sequence shown in FIG. 1 or 3(SEQ ID NO:1, transcript sequence and SEQ ID NO:3, genomic sequence), orany nucleic acid molecule that encodes the protein provided in FIG. 2,SEQ ID NO:2. A nucleic acid molecule consists of a nucleotide sequencewhen the nucleotide sequence is the complete nucleotide sequence of thenucleic acid molecule.

[0109] The present invention further provides nucleic acid moleculesthat consist essentially of the nucleotide sequence shown in FIG. 1 or 3(SEQ ID NO:1, transcript sequence and SEQ ID NO:3, genomic sequence), orany nucleic acid molecule that encodes the protein provided in FIG. 2,SEQ ID NO:2. A nucleic acid molecule consists essentially of anucleotide sequence when such a nucleotide sequence is present with onlya few additional nucleic acid residues in the final nucleic acidmolecule.

[0110] The present invention further provides nucleic acid moleculesthat comprise the nucleotide sequences shown in FIG. 1 or 3 (SEQ IDNO:1, transcript sequence and SEQ ID NO:3, genomic sequence), or anynucleic acid molecule that encodes the protein provided in FIG. 2, SEQID NO:2. A nucleic acid molecule comprises a nucleotide sequence whenthe nucleotide sequence is at least part of the final nucleotidesequence of the nucleic acid molecule. In such a fashion, the nucleicacid molecule can be only the nucleotide sequence or have additionalnucleic acid residues, such as nucleic acid residues that are naturallyassociated with it or heterologous nucleotide sequences. Such a nucleicacid molecule can have a few additional nucleotides or can comprisesseveral hundred or more additional nucleotides. A brief description ofhow various types of these nucleic acid molecules can be readilymade/isolated is provided below.

[0111] In FIGS. 1 and 3, both coding and non-coding sequences areprovided. Because of the source of the present invention, humans genomicsequence (FIG. 3) and cDNA/transcript sequences (FIG. 1), the nucleicacid molecules in the Figures will contain genomic intronic sequences,5′ and 3′ non-coding sequences, gene regulatory regions and non-codingintergenic sequences. In general such sequence features are either notedin FIGS. 1 and 3 or can readily be identified using computational toolsknown in the art. As discussed below, some of the non-coding regions,particularly gene regulatory elements such as promoters, are useful fora variety of purposes, e.g. control of heterologous gene expression,target for identifying gene activity modulating compounds, and areparticularly claimed as fragments of the genomic sequence providedherein.

[0112] The isolated nucleic acid molecules can encode the mature proteinplus additional amino or carboxyl-terminal amino acids, or amino acidsinterior to the mature peptide (when the mature form has more than onepeptide chain, for instance). Such sequences may play a role inprocessing of a protein from precursor to a mature form, facilitateprotein trafficking, prolong or shorten protein half-life or facilitatemanipulation of a protein for assay or production, among other things.As generally is the case in situ, the additional amino acids may beprocessed away from the mature protein by cellular enzymes.

[0113] As mentioned above, the isolated nucleic acid molecules include,but are not limited to, the sequence encoding the kinase peptide alone,the sequence encoding the mature peptide and additional codingsequences, such as a leader or secretory sequence (e.g., a pre-pro orpro-protein sequence), the sequence encoding the mature peptide, with orwithout the additional coding sequences, plus additional non-codingsequences, for example introns and non-coding 5′ and 3′ sequences suchas transcribed but non-translated sequences that play a role intranscription, mRNA processing (including splicing and polyadenylationsignals), ribosome binding and stability of mRNA. In addition, thenucleic acid molecule may be fused to a marker sequence encoding, forexample, a peptide that facilitates purification.

[0114] Isolated nucleic acid molecules can be in the form of RNA, suchas mRNA, or in the form DNA, including cDNA and genomic DNA obtained bycloning or produced by chemical synthetic techniques or by a combinationthereof. The nucleic acid, especially DNA, can be double-stranded orsingle-stranded. Single-stranded nucleic acid can be the coding strand(sense strand) or the non-coding strand (anti-sense strand).

[0115] The invention further provides nucleic acid molecules that encodefragments of the peptides of the present invention as well as nucleicacid molecules that encode obvious variants of the kinase proteins ofthe present invention that are described above. Such nucleic acidmolecules may be naturally occurring, such as allelic variants (samelocus), paralogs (different locus), and orthologs (different organism),or may be constructed by recombinant DNA methods or by chemicalsynthesis. Such non-naturally occurring variants may be made bymutagenesis techniques, including those applied to nucleic acidmolecules, cells, or organisms. Accordingly, as discussed above, thevariants can contain nucleotide substitutions, deletions, inversions andinsertions. Variation can occur in either or both the coding andnon-coding regions. The variations can produce both conservative andnon-conservative amino acid substitutions. The present invention furtherprovides non-coding fragments of the nucleic acid molecules provided inFIGS. 1 and 3. Preferred non-coding fragments include, but are notlimited to, promoter sequences, enhancer sequences, gene modulatingsequences and gene termination sequences. Such fragments are useful incontrolling heterologous gene expression and in developing screens toidentify gene-modulating agents. A promoter can readily be identified asbeing 5′ to the ATG start site in the genomic sequence provided in FIG.3.

[0116] A fragment comprises a contiguous nucleotide sequence greaterthan 12 or more nucleotides. Further, a fragment could at least 30, 40,50, 100, 250 or 500 nucleotides in length. The length of the fragmentwill be based on its intended use. For example, the fragment can encodeepitope bearing regions of the peptide, or can be useful as DNA probesand primers. Such fragments can be isolated using the known nucleotidesequence to synthesize an oligonucleotide probe. A labeled probe canthen be used to screen a cDNA library, genomic DNA library, or mRNA toisolate nucleic acid corresponding to the coding region. Further,primers can be used in PCR reactions to clone specific regions of gene.

[0117] A probe/primer typically comprises substantially a purifiedoligonucleotide or oligonucleotide pair. The oligonucleotide typicallycomprises a region of nucleotide sequence that hybridizes understringent conditions to at least about 12, 20, 25, 40, 50 or moreconsecutive nucleotides.

[0118] Orthologs, homologs, and allelic variants can be identified usingmethods well known in the art. As described in the Peptide Section,these variants comprise a nucleotide sequence encoding a peptide that istypically 60-70%, 70-80%, 80-90%, and more typically at least about90-95% or more homologous to the nucleotide sequence shown in the Figuresheets or a fragment of this sequence. Such nucleic acid molecules canreadily be identified as being able to hybridize under moderate tostringent conditions, to the nucleotide sequence shown in the Figuresheets or a fragment of the sequence. Allelic variants can readily bedetermined by genetic locus of the encoding gene. As indicated in FIG.3, the map position was determined to be on human chromosome 2.

[0119]FIG. 3 provides information on SNPs that have been found in thegene encoding the kinase proteins of the present invention. SNPs wereidentified at 39 different nucleotide positions, including five SNPs incoding regions, two of which (at nucleotide positions 52048 and 58826)change the encoded amino acid. The changes in the amino acid sequencethat these SNPs cause is indicated in FIG. 3 and can readily bedetermined using the universal genetic code and the protein sequenceprovided in FIG. 2 as a reference.

[0120] As used herein, the term “hybridizes under stringent conditions”is intended to describe conditions for hybridization and washing underwhich nucleotide sequences encoding a peptide at least 60-70% homologousto each other typically remain hybridized to each other. The conditionscan be such that sequences at least about 60%, at least about 70%, or atleast about 80% or more homologous to each other typically remainhybridized to each other. Such stringent conditions are known to thoseskilled in the art and can be found in Current Protocols in MolecularBiology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. One example ofstringent hybridization conditions are hybridization in 6× sodiumchloride/sodium citrate (SSC) at about 45C., followed by one or morewashes in 0.2×SSC, 0.1% SDS at 50-65C. Examples of moderate to lowstringency hybridization conditions are well known in the art.

[0121] Nucleic Acid Molecule Uses

[0122] The nucleic acid molecules of the present invention are usefulfor probes, primers, chemical intermediates, and in biological assays.The nucleic acid molecules are useful as a hybridization probe formessenger RNA, transcript/cDNA and genomic DNA to isolate full-lengthcDNA and genomic clones encoding the peptide described in FIG. 2 and toisolate cDNA and genomic clones that correspond to variants (alleles,orthologs, etc.) producing the same or related peptides shown in FIG. 2.As illustrated in FIG. 3, SNPs were identified at 39 differentnucleotide positions, including 2 non-synonymous coding SNPs.

[0123] The probe can correspond to any sequence along the entire lengthof the nucleic acid molecules provided in the Figures. Accordingly, itcould be derived from 5′ noncoding regions, the coding region, and 3′noncoding regions. However, as discussed, fragments are not to beconstrued as encompassing fragments disclosed prior to the presentinvention.

[0124] The nucleic acid molecules are also useful as primers for PCR toamplify any given region of a nucleic acid molecule and are useful tosynthesize antisense molecules of desired length and sequence.

[0125] The nucleic acid molecules are also useful for constructingrecombinant vectors. Such vectors include expression vectors thatexpress a portion of, or all of, the peptide sequences. Vectors alsoinclude insertion vectors, used to integrate into another nucleic acidmolecule sequence, such as into the cellular genome, to alter in situexpression of a gene and/or gene product. For example, an endogenouscoding sequence can be replaced via homologous recombination with all orpart of the coding region containing one or more specifically introducedmutations.

[0126] The nucleic acid molecules are also useful for expressingantigenic portions of the proteins.

[0127] The nucleic acid molecules are also useful as probes fordetermining the chromosomal positions of the nucleic acid molecules bymeans of in situ hybridization methods. As indicated in FIG. 3, the mapposition was determined to be on human chromosome 2.

[0128] The nucleic acid molecules are also useful in making vectorscontaining the gene regulatory regions of the nucleic acid molecules ofthe present invention.

[0129] The nucleic acid molecules are also useful for designingribozymes corresponding to all, or a part, of the mRNA produced from thenucleic acid molecules described herein.

[0130] The nucleic acid molecules are also useful for making vectorsthat express part, or all, of the peptides.

[0131] The nucleic acid molecules are also useful for constructing hostcells expressing a part, or all, of the nucleic acid molecules andpeptides.

[0132] The nucleic acid molecules are also useful for constructingtransgenic animals expressing all, or a part, of the nucleic acidmolecules and peptides.

[0133] The nucleic acid molecules are also useful as hybridizationprobes for determining the presence, level, form and distribution ofnucleic acid expression. Experimental data as provided in FIG. 1indicates that kinase proteins of the present invention are expressed inadult and fetal brain (including astrocytoma and neuroblastoma cells),lung/spleen, and squamous cell carcinoma (skin), as indicated by virtualnorthern blot analysis. Accordingly, the probes can be used to detectthe presence of, or to determine levels of, a specific nucleic acidmolecule in cells, tissues, and in organisms. The nucleic acid whoselevel is determined can be DNA or RNA. Accordingly, probes correspondingto the peptides described herein can be used to assess expression and/orgene copy number in a given cell, tissue, or organism. These uses arerelevant for diagnosis of disorders involving an increase or decrease inkinase protein expression relative to normal results.

[0134] In vitro techniques for detection of mRNA include Northernhybridizations and in situ hybridizations. In vitro techniques fordetecting DNA includes Southern hybridizations and in situhybridization.

[0135] Probes can be used as a part of a diagnostic test kit foridentifying cells or tissues that express a kinase protein, such as bymeasuring a level of a kinase-encoding nucleic acid in a sample of cellsfrom a subject e.g., mRNA or genomic DNA, or determining if a kinasegene has been mutated. Experimental data as provided in FIG. 1 indicatesthat kinase proteins of the. present invention are expressed in adultand fetal brain (including astrocytoma and neuroblastoma cells),lung/spleen, and squamous cell carcinoma (skin), as indicated by virtualnorthern blot analysis.

[0136] Nucleic acid expression assays are useful for drug screening toidentify compounds that modulate kinase nucleic acid expression.

[0137] The invention thus provides a method for identifying a compoundthat can be used to treat a disorder associated with nucleic acidexpression of the kinase gene, particularly biological and pathologicalprocesses that are mediated by the kinase in cells and tissues thatexpress it. Experimental data as provided in FIG. 1 indicates expressionin adult and fetal brain (including astrocytoma and neuroblastomacells), lung/spleen, and squamous cell carcinoma (skin). The methodtypically includes assaying the ability of the compound to modulate theexpression of the kinase nucleic acid and thus identifying a compoundthat can be used to treat a disorder characterized by undesired kinasenucleic acid expression. The assays can be performed in cell-based andcell-free systems. Cell-based assays include cells naturally expressingthe kinase nucleic acid or recombinant cells genetically engineered toexpress specific nucleic acid sequences.

[0138] The assay for kinase nucleic acid expression can involve directassay of nucleic acid levels, such as mRNA levels, or on collateralcompounds involved in the signal pathway. Further, the expression ofgenes that are up- or down-regulated in response to the kinase proteinsignal pathway can also be assayed. In this embodiment the regulatoryregions of these genes can be operably linked to a reporter gene such asluciferase.

[0139] Thus, modulators of kinase gene expression can be identified in amethod wherein a cell is contacted with a candidate compound and theexpression of mRNA determined. The level of expression of kinase mRNA inthe presence of the candidate compound is compared to the level ofexpression of kinase mRNA in the absence of the candidate compound. Thecandidate compound can then be identified as a modulator of nucleic acidexpression based on this comparison and be used, for example to treat adisorder characterized by aberrant nucleic acid expression. Whenexpression of mRNA is statistically significantly greater in thepresence of the candidate compound than in its absence, the candidatecompound is identified as a stimulator of nucleic acid expression. Whennucleic acid expression is statistically significantly less in thepresence of the candidate compound than in its absence, the candidatecompound is identified as an inhibitor of nucleic acid expression.

[0140] The invention further provides methods of treatment, with thenucleic acid as a target, using a compound identified through drugscreening as a gene modulator to modulate kinase nucleic acid expressionin cells and tissues that express the kinase. Experimental data asprovided in FIG. 1 indicates that kinase proteins of the presentinvention are expressed in adult and fetal brain (including astrocytomaand neuroblastoma cells), lung/spleen, and squamous cell carcinoma(skin), as indicated by virtual northern blot analysis. Modulationincludes both up-regulation (i.e. activation or agonization) ordown-regulation (suppression or antagonization) or nucleic acidexpression.

[0141] Alternatively, a modulator for kinase nucleic acid expression canbe a small molecule or drug identified using the screening assaysdescribed herein as long as the drug or small molecule inhibits thekinase nucleic acid expression in the cells and tissues that express theprotein. Experimental data as provided in FIG. 1 indicates expression inadult and fetal brain (including astrocytoma and neuroblastoma cells),lung/spleen, and squamous cell carcinoma (skin).

[0142] The nucleic acid molecules are also useful for monitoring theeffectiveness of modulating compounds on the expression or activity ofthe kinase gene in clinical trials or in a treatment regimen. Thus, thegene expression pattern can serve as a barometer for the continuingeffectiveness of treatment with the compound, particularly withcompounds to which a patient can develop resistance. The gene expressionpattern can also serve as a marker indicative of a physiologicalresponse of the affected cells to the compound. Accordingly, suchmonitoring would allow either increased administration of the compoundor the administration of alternative compounds to which the patient hasnot become resistant. Similarly, if the level of nucleic acid expressionfalls below a desirable level, administration of the compound could becommensurately decreased.

[0143] The nucleic acid molecules are also useful in diagnostic assaysfor qualitative changes in kinase nucleic acid expression, andparticularly in qualitative changes that lead to pathology. The nucleicacid molecules can be used to detect mutations in kinase genes and geneexpression products such as mRNA. The nucleic acid molecules can be usedas hybridization probes to detect naturally occurring genetic mutationsin the kinase gene and thereby to determine whether a subject with themutation is at risk for a disorder caused by the mutation. Mutationsinclude deletion, addition, or substitution of one or more nucleotidesin the gene, chromosomal rearrangement, such as inversion ortransposition, modification of genomic DNA, such as aberrant methylationpatterns or changes in gene copy number, such as amplification.Detection of a mutated form of the kinase gene associated with adysfunction provides a diagnostic tool for an active disease orsusceptibility to disease when the disease results from overexpression,underexpression, or altered expression of a kinase protein.

[0144] Individuals carrying mutations in the kinase gene can be detectedat the nucleic acid level by a variety of techniques. FIG. 3 providesinformation on SNPs that have been found in the gene encoding the kinaseproteins of the present invention. SNPs were identified at 39 differentnucleotide positions, including five SNPs in coding regions, two ofwhich (at nucleotide positions 52048 and 58826) change the encoded aminoacid. The changes in the amino acid sequence that these SNPs cause isindicated in FIG. 3 and can readily be determined using the universalgenetic code and the protein sequence provided in FIG. 2 as a reference.As indicated in FIG. 3, the map position was determined to be on humanchromosome 2. Genomic DNA can be analyzed directly or can be amplifiedby using PCR prior to analysis. RNA or cDNA can be used in the same way.In some uses, detection of the mutation involves the use of aprobe/primer in a polymerase chain reaction (PCR) (see, e.g. U.S. Pat.Nos. 4,683,195 and 4,683,202), such as anchor PCR or RACE PCR, or,alternatively, in a ligation chain reaction (LCR) (see, e.g., Landegranet al., Science 241:1077-1080 (1988); and Nakazawa et al., PNAS91:360-364 (1994)), the latter of which can be particularly useful fordetecting point mutations in the gene (see Abravaya et al., NucleicAcids Res. 23:675-682 (1995)). This method can include the steps ofcollecting a sample of cells from a patient, isolating nucleic acid(e.g., genomic, mRNA or both) from the cells of the sample, contactingthe nucleic acid sample with one or more primers which specificallyhybridize to a gene under conditions such that hybridization andamplification of the gene (if present) occurs, and detecting thepresence or absence of an amplification product, or detecting the sizeof the amplification product and comparing the length to a controlsample. Deletions and insertions can be detected by a change in size ofthe amplified product compared to the normal genotype. Point mutationscan be identified by hybridizing amplified DNA to normal RNA orantisense DNA sequences.

[0145] Alternatively, mutations in a kinase gene can be directlyidentified, for example, by alterations in restriction enzyme digestionpatterns determined by gel electrophoresis.

[0146] Further, sequence-specific ribozymes (U.S. Pat. No. 5,498,531)can be used to score for the presence of specific mutations bydevelopment or loss of a ribozyme cleavage site. Perfectly matchedsequences can be distinguished from mismatched sequences by nucleasecleavage digestion assays or by differences in melting temperature.

[0147] Sequence changes at specific locations can also be assessed bynuclease protection assays such as RNase and S1 protection or thechemical cleavage method. Furthermore, sequence differences between amutant kinase gene and a wild-type gene can be determined by direct DNAsequencing. A variety of automated sequencing procedures can be utilizedwhen performing the diagnostic assays (Naeve, C. W., (1995)Biotechniques 19:448), including sequencing by mass spectrometry (see,e.g., PCT International Publication No. WO 94/16101; Cohen et al., Adv.Chromatogr. 36:127-162 (1996); and Griffin et al., Appl. Biochem.Biotechnol., 38:147-159 (1993)).

[0148] Other methods for detecting mutations in the gene include methodsin which protection from cleavage agents is used to detect mismatchedbases in RNA/RNA or RNA/DNA duplexes (Myers et al., Science 230:1242(1985)); Cotton et al., PNAS 85:4397(1988); Saleeba et al., Meth.Enzymol. 217:286-295 (1992)), electrophoretic mobility of mutant andwild type nucleic acid is compared (Orita et al., PNAS 86:2766 (1989);Cotton et al., Mutat. Res. 285:125-144 (1993); and Hayashi et al.,Genet. Anal. Tech. Appl. 9:73-79 (1992)), and movement of mutant orwild-type fragments in polyacrylamide gels containing a gradient ofdenaturant is assayed using denaturing gradient gel electrophoresis(Myers et al., Nature 313:495 (1985)). Examples of other techniques fordetecting point mutations include selective oligonucleotidehybridization, selective amplification, and selective primer extension.

[0149] The nucleic acid molecules are also useful for testing anindividual for a genotype that while not necessarily causing thedisease, nevertheless affects the treatment modality. Thus, the nucleicacid molecules can be used to study the relationship between anindividual's genotype and the individual's response to a compound usedfor treatment (pharmacogenomic relationship). Accordingly, the nucleicacid molecules described herein can be used to assess the mutationcontent of the kinase gene in an individual in order to select anappropriate compound or dosage regimen for treatment. FIG. 3 providesinformation on SNPs that have been found in the gene encoding the kinaseproteins of the present invention. SNPs were identified at 39 differentnucleotide positions, including five SNPs in coding regions, two ofwhich (at nucleotide positions 52048 and 58826) change the encoded aminoacid. The changes in the amino acid sequence that these SNPs cause isindicated in FIG. 3 and can readily be determined using the universalgenetic code and the protein sequence provided in FIG. 2 as a reference.

[0150] Thus nucleic acid molecules displaying genetic variations thataffect treatment provide a diagnostic target that can be used to tailortreatment in an individual. Accordingly, the production of recombinantcells and animals containing these polymorphisms allow effectiveclinical design of treatment compounds and dosage regimens.

[0151] The nucleic acid molecules are thus useful as antisenseconstructs to control kinase gene expression in cells, tissues, andorganisms. A DNA antisense nucleic acid molecule is designed to becomplementary to a region of the gene involved in transcription,preventing transcription and hence production of kinase protein. Anantisense RNA or DNA nucleic acid molecule would hybridize to the mRNAand thus block translation of mRNA into kinase protein.

[0152] Alternatively, a class of antisense molecules can be used toinactivate mRNA in order to decrease expression of kinase nucleic acid.Accordingly, these molecules can treat a disorder characterized byabnormal or undesired kinase nucleic acid expression. This techniqueinvolves cleavage by means of ribozymes containing nucleotide sequencescomplementary to one or more regions in the mRNA that attenuate theability of the mRNA to be translated. Possible regions include codingregions and particularly coding regions corresponding to the catalyticand other functional activities of the kinase protein, such as substratebinding.

[0153] The nucleic acid molecules also provide vectors for gene therapyin patients containing cells that are aberrant in kinase geneexpression. Thus, recombinant cells, which include the patient's cellsthat have been engineered ex vivo and returned to the patient, areintroduced into an individual where the cells produce the desired kinaseprotein to treat the individual.

[0154] The invention also encompasses kits for detecting the presence ofa kinase nucleic acid in a biological sample. Experimental data asprovided in FIG. 1 indicates that kinase proteins of the presentinvention are expressed in adult and fetal brain (including astrocytomaand neuroblastoma cells), lung/spleen, and squamous cell carcinoma(skin), as indicated by virtual northern blot analysis. For example, thekit can comprise reagents such as a labeled or labelable nucleic acid oragent capable of detecting kinase nucleic acid in a biological sample;means for determining the amount of kinase nucleic acid in the sample;and means for comparing the amount of kinase nucleic acid in the samplewith a standard. The compound or agent can be packaged in a suitablecontainer. The kit can further comprise instructions for using the kitto detect kinase protein mRNA or DNA.

[0155] Nucleic Acid Arrays

[0156] The present invention further provides nucleic acid detectionkits, such as arrays or microarrays of nucleic acid molecules that arebased on the sequence information provided in FIGS. 1 and 3 (SEQ ID NOS:1and 3).

[0157] As used herein “Arrays” or “Microarrays” refers to an array ofdistinct polynucleotides or oligonucleotides synthesized on a substrate,such as paper, nylon or other type of membrane, filter, chip, glassslide, or any other suitable solid support. In one embodiment, themicroarray is prepared and used according to the methods described inU.S. Pat. No. 5,837,832, Chee et al., PCT application WO95/11995 (Cheeet al.), Lockhart, D. J. et al. (1996; Nat. Biotech. 14: 1675-1680) andSchena, M. et al. (1996; Proc. Natl. Acad. Sci. 93: 10614-10619), all ofwhich are incorporated herein in their entirety by reference. In otherembodiments, such arrays are produced by the methods described by Brownet al., U.S. Pat. No. 5,807,522.

[0158] The microarray or detection kit is preferably composed of a largenumber of unique, single-stranded nucleic acid sequences, usually eithersynthetic antisense oligonucleotides or fragments of cDNAs, fixed to asolid support. The oligonucleotides are preferably about 6-60nucleotides in length, more preferably 15-30 nucleotides in length, andmost preferably about 20-25 nucleotides in length. For a certain type ofmicroarray or detection kit, it may be preferable to useoligonucleotides that are only 7-20 nucleotides in length. Themicroarray or detection kit may contain oligonucleotides that cover theknown 5′, or 3′, sequence, sequential oligonucleotides which cover thefull length sequence; or unique oligonucleotides selected fromparticular areas along the length of the sequence. Polynucleotides usedin the microarray or detection kit may be oligonucleotides that arespecific to a gene or genes of interest.

[0159] In order to produce oligonucleotides to a known sequence for amicroarray or detection kit, the gene(s) of interest (or an ORFidentified from the contigs of the present invention) is typicallyexamined using a computer algorithm which starts at the 5′ or at the 3′end of the nucleotide sequence. Typical algorithms will then identifyoligomers of defined length that are unique to the gene, have a GCcontent within a range suitable for hybridization, and lack predictedsecondary structure that may interfere with hybridization. In certainsituations it may be appropriate to use pairs of oligonucleotides on amicroarray or detection kit. The “pairs” will be identical, except forone nucleotide that preferably is located in the center of the sequence.The second oligonucleotide in the pair (mismatched by one) serves as acontrol. The number of oligonucleotide pairs may range from two to onemillion. The oligomers are synthesized at designated areas on asubstrate using a light-directed chemical process. The substrate may bepaper, nylon or other type of membrane, filter, chip, glass slide or anyother suitable solid support.

[0160] In another aspect, an oligonucleotide may be synthesized on thesurface of the substrate by using a chemical coupling procedure and anink jet application apparatus, as described in PCT applicationWO95/251116 (Baldeschweiler et al.) which is incorporated herein in itsentirety by reference. In another aspect, a “gridded” array analogous toa dot (or slot) blot may be used to arrange and link cDNA fragments oroligonucleotides to the surface of a substrate using a vacuum system,thermal, UV, mechanical or chemical bonding procedures. An array, suchas those described above, may be produced by hand or by using availabledevices (slot blot or dot blot apparatus), materials (any suitable solidsupport), and machines (including robotic instruments), and may contain8, 24, 96, 384, 1536, 6144 or more oligonucleotides, or any other numberbetween two and one million which lends itself to the efficient use ofcommercially available instrumentation.

[0161] In order to conduct sample analysis using a microarray ordetection kit, the RNA or DNA from a biological sample is made intohybridization probes. The mRNA is isolated, and cDNA is produced andused as a template to make antisense RNA (aRNA). The aRNA is amplifiedin the presence of fluorescent nucleotides, and labeled probes areincubated with the microarray or detection kit so that the probesequences hybridize to complementary oligonucleotides of the microarrayor detection kit. Incubation conditions are adjusted so thathybridization occurs with precise complementary matches or with variousdegrees of less complementarity. After removal of nonhybridized probes,a scanner is used to determine the levels and patterns of fluorescence.The scanned images are examined to determine degree of complementarityand the relative abundance of each oligonucleotide sequence on themicroarray or detection kit. The biological samples may be obtained fromany bodily fluids (such as blood, urine, saliva, phlegm, gastric juices,etc.), cultured cells, biopsies, or other tissue preparations. Adetection system may be used to measure the absence, presence, andamount of hybridization for all of the distinct sequencessimultaneously. This data may be used for large-scale correlationstudies on the sequences, expression patterns, mutations, variants, orpolymorphisms among samples.

[0162] Using such arrays, the present invention provides methods toidentify the expression of the kinase proteins/peptides of the presentinvention. In detail, such methods comprise incubating a test samplewith one or more nucleic acid molecules and assaying for binding of thenucleic acid molecule with components within the test sample. Suchassays will typically involve arrays comprising many genes, at least oneof which is a gene of the present invention and or alleles of the kinasegene of the present invention. FIG. 3 provides information on SNPs thathave been found in the gene encoding the kinase proteins of the presentinvention. SNPs were identified at 39. different nucleotide positions,including five SNPs in coding regions, two of which (at nucleotidepositions 52048 and 58826) change the encoded amino acid. The changes inthe amino acid sequence that these SNPs cause is indicated in FIG. 3 andcan readily be determined using the universal genetic code and theprotein sequence provided in FIG. 2 as a reference.

[0163] Conditions for incubating a nucleic acid molecule with a testsample vary. Incubation conditions depend on the format employed in theassay, the detection methods employed, and the type and nature of thenucleic acid molecule used in the assay. One skilled in the art willrecognize that any one of the commonly available hybridization,amplification or array assay formats can readily be adapted to employthe novel fragments of the Human genome disclosed herein. Examples ofsuch assays can be found in Chard, T, An Introduction toRadioimmunoassay and Related Techniques, Elsevier Science Publishers,Amsterdam, The Netherlands (1986); Bullock, G. R. et al., Techniques inImmunocytochemistry, Academic Press, Orlando, Fla. Vol. 1 (1982), Vol. 2(1983), Vol. 3 (1985); Tijssen, P., Practice and Theory of EnzymeImmunoassays: Laboratory Techniques in Biochemistry and MolecularBiology, Elsevier Science Publishers, Amsterdam, The Netherlands (1985).

[0164] The test samples of the present invention include cells, proteinor membrane extracts of cells. The test sample used in theabove-described method will vary based on the assay format, nature ofthe detection method and the tissues, cells or extracts used as thesample to be assayed. Methods for preparing nucleic acid extracts or ofcells are well known in the art and can be readily be adapted in orderto obtain a sample that is compatible with the system utilized.

[0165] In another embodiment of the present invention, kits are providedwhich contain the necessary reagents to carry out the assays of thepresent invention.

[0166] Specifically, the invention provides a compartmentalized kit toreceive, in close confinement, one or more containers which comprises:(a) a first container comprising one of the nucleic acid molecules thatcan bind to a fragment of the Human genome disclosed herein; and (b) oneor more other containers comprising one or more of the following: washreagents, reagents capable of detecting presence of a bound nucleicacid.

[0167] In detail, a compartmentalized kit includes any kit in whichreagents are contained in separate containers. Such containers includesmall glass containers, plastic containers, strips of plastic, glass orpaper, or arraying material such as silica. Such containers allows oneto efficiently transfer reagents from one compartment to anothercompartment such that the samples and reagents are notcross-contaminated, and the agents or solutions of each container can beadded in a quantitative fashion from one compartment to another. Suchcontainers will include a container which will accept the test sample, acontainer which contains the nucleic acid probe, containers whichcontain wash reagents (such as phosphate buffered saline, Tris-buffers,etc.), and containers which contain the reagents used to detect thebound probe. One skilled in the art will readily recognize that thepreviously unidentified kinase gene of the present invention can beroutinely identified using the sequence information disclosed herein canbe readily incorporated into one of the established kit formats whichare well known in the art, particularly expression arrays.

[0168] Vectors/Host Cells

[0169] The invention also provides vectors containing the nucleic acidmolecules described herein. The term “vector” refers to a vehicle,preferably a nucleic acid molecule, which can transport the nucleic acidmolecules. When the vector is a nucleic acid molecule, the nucleic acidmolecules are covalently linked to the vector nucleic acid. With thisaspect of the invention, the vector includes a plasmid, single or doublestranded phage, a single or double stranded RNA or DNA viral vector, orartificial chromosome, such as a BAC, PAC, YAC, OR MAC.

[0170] A vector can be maintained in the host cell as anextrachromosomal element where it replicates and produces additionalcopies of the nucleic acid molecules. Alternatively, the vector mayintegrate into the host cell genome and produce additional copies of thenucleic acid molecules when the host cell replicates.

[0171] The invention provides vectors for the maintenance (cloningvectors) or vectors for expression (expression vectors) of the nucleicacid molecules. The vectors can function in prokaryotic or eukaryoticcells or in both (shuttle vectors).

[0172] Expression vectors contain cis-acting regulatory regions that areoperably linked in the vector to the nucleic acid molecules such thattranscription of the nucleic acid molecules is allowed in a host cell.The nucleic acid molecules can be introduced into the host cell with aseparate nucleic acid molecule capable of affecting transcription. Thus,the second nucleic acid molecule may provide a trans-acting factorinteracting with the cis-regulatory control region to allowtranscription of the nucleic acid molecules from the vector.Alternatively, a trans-acting factor may be supplied by the host cell.Finally, a trans-acting factor can be produced from the vector itself.It is understood, however, that in some embodiments, transcriptionand/or translation of the nucleic acid molecules can occur in acell-free system.

[0173] The regulatory sequence to which the nucleic acid moleculesdescribed herein can be operably linked include promoters for directingmRNA transcription. These include, but are not limited to, the leftpromoter from bacteriophage λ, the lac, TRP, and TAC promoters from E.coli, the early and late promoters from SV40, the CMV immediate earlypromoter, the adenovirus early and late promoters, and retroviruslong-terminal repeats.

[0174] In addition to control regions that promote transcription,expression vectors may also include regions that modulate transcription,such as repressor binding sites and enhancers. Examples include the SV40enhancer, the cytomegalovirus immediate early enhancer, polyomaenhancer, adenovirus enhancers, and retrovirus LTR enhancers.

[0175] In addition to containing sites for transcription initiation andcontrol, expression vectors can also contain sequences necessary fortranscription termination and, in the transcribed region a ribosomebinding site for translation. Other regulatory control elements forexpression include initiation and termination codons as well aspolyadenylation signals. The person of ordinary skill in the art wouldbe aware of the numerous regulatory sequences that are useful inexpression vectors. Such regulatory sequences are described, forexample, in Sambrook et al., Molecular Cloning: A Laboratory Manual.2nd. ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.,(1989).

[0176] A variety of expression vectors can be used to express a nucleicacid molecule. Such vectors include chromosomal, episomal, andvirus-derived vectors, for example vectors derived from bacterialplasmids, from bacteriophage, from yeast episomes, from yeastchromosomal elements, including yeast artificial chromosomes, fromviruses such as baculoviruses, papovaviruses such as SV40, Vacciniaviruses, adenoviruses, poxviruses, pseudorabies viruses, andretroviruses. Vectors may also be derived from combinations of thesesources such as those derived from plasmid and bacteriophage geneticelements, e.g. cosmids and phagemids. Appropriate cloning and expressionvectors for prokaryotic and eukaryotic hosts are described in Sambrooket al., Molecular Cloning:. A Laboratory Manual. 2nd. ed., Cold SpringHarbor Laboratory Press, Cold Spring Harbor, N.Y., (1989).

[0177] The regulatory sequence may provide constitutive expression inone or more host cells (i.e. tissue specific) or may provide forinducible expression in one or more cell types such as by temperature,nutrient additive, or exogenous factor such as a hormone or otherligand. A variety of vectors providing for constitutive and inducibleexpression in prokaryotic and eukaryotic hosts are well known to thoseof ordinary skill in the art.

[0178] The nucleic acid molecules can be inserted into the vectornucleic acid by well-known methodology. Generally, the DNA sequence thatwill ultimately be expressed is joined to an expression vector bycleaving the DNA sequence and the expression vector with one or morerestriction enzymes and then ligating the fragments together. Proceduresfor restriction enzyme digestion and ligation are well known to those ofordinary skill in the art.

[0179] The vector containing the appropriate nucleic acid molecule canbe introduced into an appropriate host cell for propagation orexpression using well-known techniques. Bacterial cells include, but arenot limited to, E. coli, Streptomyces, and Salmonella typhimurium.Eukaryotic cells include, but are not limited to, yeast, insect cellssuch as Drosophila, animal cells such as COS and CHO cells, and plantcells.

[0180] As described herein, it may be desirable to express the peptideas a fusion protein. Accordingly, the invention provides fusion vectorsthat allow for the production of the peptides. Fusion vectors canincrease the expression of a recombinant protein, increase thesolubility of the recombinant protein, and aid in the purification ofthe protein by acting for example as a ligand for affinity purification.A proteolytic cleavage site may be introduced at the junction of thefusion moiety so that the desired peptide can ultimately be separatedfrom the fusion moiety. Proteolytic enzymes include, but are not limitedto, factor Xa, thrombin, and enterokinase. Typical fusion expressionvectors include pGEX (Smith et al., Gene 67:31-40 (1988)), pMAL (NewEngland Biolabs, Beverly, Mass.) and pRIT5 (Pharmacia, Piscataway, N.J.)which fuse glutathione S-transferase (GST), maltose E binding protein,or protein A, respectively, to the target recombinant protein. Examplesof suitable inducible non-fusion E. coli expression vectors include pTrc(Amann et al., Gene 69:301-315 (1988)) and pET 11d (Studier et al., GeneExpression Technology: Methods in Enzymology 185:60-89 (1990)).

[0181] Recombinant protein expression can be maximized in host bacteriaby providing a genetic background wherein the host cell has an impairedcapacity to proteolytically cleave the recombinant protein. (Gottesman,S., Gene Expression Technology: Methods in Enzymology 185, AcademicPress, San Diego, Calif. (1990) 119-128). Alternatively, the sequence ofthe nucleic acid molecule of interest can be altered to providepreferential codon usage for a specific host cell, for example E. coli.(Wada et al., Nucleic Acids Res. 20:2111-2118 (1992)).

[0182] The nucleic acid molecules can also be expressed by expressionvectors that are operative in yeast. Examples of vectors for expressionin yeast e.g., S. cerevisiae include pYepSec1 (Baldari, et-al., EMBO J.6:229-234 (1987)), pMFa (Kujan et al., Cell 30:933-943(1982)), pJRY88(Schultz et al., Gene 54:113-123 (1987)), and pYES2 (InvitrogenCorporation, San Diego, Calif.).

[0183] The nucleic acid molecules can also be expressed in insect cellsusing, for example, baculovirus expression vectors. Baculovirus vectorsavailable for expression of proteins in cultured insect cells (e.g., Sf9cells) include the pAc series (Smith et al., Mol. Cell Biol. 3:2156-2165(1983)) and the pVL series (Lucklow et al., Virology 170:31-39 (1989)).

[0184] In certain embodiments of the invention, the nucleic acidmolecules described herein are expressed in mammalian cells usingmammalian expression vectors. Examples of mammalian expression vectorsinclude pCDM8 (Seed, B. Nature 329:840(1987)) and pMT2PC (Kaufman etal., EMBO J. 6:187-195 (1987)).

[0185] The expression vectors listed herein are provided by way ofexample only of the well-known vectors available to those of ordinaryskill in the art that would be useful to express the nucleic acidmolecules. The person of ordinary skill in the art would be aware ofother vectors suitable for maintenance propagation or expression of thenucleic acid molecules described herein. These are found for example inSambrook, J., Fritsh, E. F., and Maniatis, T. Molecular Cloning: ALaboratory Manual. 2nd., ed., Cold Spring Harbor Laboratory, Cold SpringHarbor Laboratory Press, Cold Spring Harbor, N.Y., 1989.

[0186] The invention also encompasses vectors in which the nucleic acidsequences described herein are cloned into the vector in reverseorientation, but operably linked to a regulatory sequence that permitstranscription of antisense RNA. Thus, an antisense transcript can beproduced to all, or to a portion, of the nucleic acid molecule sequencesdescribed herein, including both coding and non-coding regions.Expression of this antisense RNA is subject to each of the parametersdescribed above in relation to expression of the sense RNA (regulatorysequences, constitutive or inducible expression, tissue-specificexpression).

[0187] The invention also relates to recombinant host cells containingthe vectors described herein. Host cells therefore include prokaryoticcells, lower eukaryotic cells such as yeast, other eukaryotic cells suchas insect cells, and higher eukaryotic cells such as mammalian cells.

[0188] The recombinant host cells are prepared by introducing the vectorconstructs described herein into the cells by techniques readilyavailable to the person of ordinary skill in the art. These include, butare not limited to, calcium phosphate transfection,DEAE-dextran-mediated transfection, cationic lipid-mediatedtransfection, electroporation, transduction, infection, lipofection, andother techniques such as those found in Sambrook, et al. (MolecularCloning: A Laboratory Manual. 2nd, ed., Cold Spring Harbor Laboratory,Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989).

[0189] Host cells can contain more than one vector. Thus, differentnucleotide sequences can be introduced on different vectors of the samecell. Similarly, the nucleic acid molecules can be introduced eitheralone or with other nucleic acid molecules that are not related to thenucleic acid molecules such as those providing trans-acting factors forexpression vectors. When more than one vector is introduced into a cell,the vectors can be introduced independently, co-introduced or joined tothe nucleic acid molecule vector.

[0190] In the case of bacteriophage and viral vectors, these can beintroduced into cells as packaged or encapsulated virus by standardprocedures for infection and transduction. Viral vectors can bereplication-competent or replication-defective. In the case in whichviral replication is defective, replication will occur in host cellsproviding functions that complement the defects.

[0191] Vectors generally include selectable markers that enable theselection of the subpopulation of cells that contain the recombinantvector constructs. The marker can be contained in the same vector thatcontains the nucleic acid molecules described herein or may be on aseparate vector. Markers include tetracycline or ampicillin-resistancegenes for prokaryotic host cells and dihydrofolate reductase or neomycinresistance for eukaryotic host cells. However, any marker that providesselection for a phenotypic trait will be effective.

[0192] While the mature proteins can be produced in bacteria, yeast,mammalian cells, and other cells under the control of the appropriateregulatory sequences, cell- free transcription and translation systemscan also be used to produce these proteins using RNA derived from theDNA constructs described herein.

[0193] Where secretion of the peptide is desired, which is difficult toachieve with multi-transmembrane domain containing proteins such askinases, appropriate secretion signals are incorporated into the vector.The signal sequence can be endogenous to the peptides or heterologous tothese peptides.

[0194] Where the peptide is not secreted into the medium, which istypically the case with kinases, the protein can be isolated from thehost cell by standard disruption procedures, including freeze thaw,sonication, mechanical disruption, use of lysing agents and the like.The peptide can then be recovered and purified by well-knownpurification methods including ammonium sulfate precipitation, acidextraction, anion or cationic exchange chromatography, phosphocellulosechromatography, hydrophobic-interaction chromatography, affinitychromatography, hydroxylapatite chromatography, lectin chromatography,or high performance liquid chromatography.

[0195] It is also understood that depending upon the host cell inrecombinant production of the peptides described herein, the peptidescan have various glycosylation patterns, depending upon the cell, ormaybe non-glycosylated as when produced in bacteria. In addition, thepeptides may include an initial modified methionine in some cases as aresult of a host-mediated process.

[0196] Uses of Vectors and Host Cells

[0197] The recombinant host cells expressing the peptides describedherein have a variety of uses. First, the cells are useful for producinga kinase protein or peptide that can be further purified to producedesired amounts of kinase protein or fragments. Thus, host cellscontaining expression vectors are useful for peptide production.

[0198] Host cells are also useful for conducting cell-based assaysinvolving the kinase protein or kinase protein fragments, such as thosedescribed above as well as other formats known in the art. Thus, arecombinant host cell expressing a native kinase protein is useful forassaying compounds that stimulate or inhibit kinase protein function.

[0199] Host cells are also useful for identifying kinase protein mutantsin which these functions are affected. If the mutants naturally occurand give rise to a pathology, host cells containing the mutations areuseful to assay compounds that have a desired effect on the mutantkinase protein (for example, stimulating or inhibiting function) whichmay not be indicated by their effect on the native kinase protein.

[0200] Genetically engineered host cells can be further used to producenon-human transgenic animals. A transgenic animal is preferably amammal, for example a rodent, such as a rat or mouse, in which one ormore of the cells of the animal include a transgene. A transgene isexogenous DNA which is integrated into the genome of a cell from which atransgenic animal develops and which remains in the genome of the matureanimal in one or more cell types or tissues of the transgenic animal.These animals are useful for studying the function of a kinase proteinand identifying and evaluating modulators of kinase protein activity.Other examples of transgenic animals include non-human primates, sheep,dogs, cows, goats, chickens, and amphibians.

[0201] A transgenic animal can be produced by introducing nucleic acidinto the male pronuclei of a fertilized oocyte, e.g., by microinjection,retroviral infection, and allowing the oocyte to develop in apseudopregnant female foster animal. Any of the kinase proteinnucleotide sequences can be introduced as a transgene into the genome ofa non-human animal, such as a mouse.

[0202] Any of the regulatory or other sequences useful in expressionvectors can form part of the transgenic sequence. This includes intronicsequences and polyadenylation signals, if not already included. Atissue-specific regulatory sequence(s) can be operably linked to thetransgene to direct expression of the kinase protein to particularcells.

[0203] Methods for generating transgenic animals via embryo manipulationand microinjection, particularly animals such as mice, have becomeconventional in the art and are described, for example, in U.S. Pat.Nos. 4,736,866 and 4,870,009, both by Leder et al., U.S. Pat. No.4,873,191 by Wagner et al. and in Hogan, B., Manipulating the MouseEmbryo, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.,1986). Similar methods are used for production of other transgenicanimals. A transgenic founder animal can be identified based upon thepresence of the transgene in its genome and/or expression of transgenicmRNA in tissues or cells of the animals. A transgenic founder animal canthen be used to breed additional animals carrying the transgene.Moreover, transgenic animals carrying a transgene can further be bred toother transgenic animals carrying other transgenes. A transgenic animalalso includes animals in which the entire animal or tissues in theanimal have been produced using the homologously recombinant host cellsdescribed herein.

[0204] In another embodiment, transgenic non-human animals can beproduced which contain selected systems that allow for regulatedexpression of the transgene. One example of such a system is thecre/loxP recombinase system of bacteriophage P1. For a description ofthe cre/loxP recombinase system, see, e.g., Lakso et al.PNAS89:6232-6236 (1992). Another example of a recombinase system is the FLPrecombinase system of S. cerevisiae (O'Gorman et al. Science251:1351-1355 (1991). If a cre/loxP recombinase system is used toregulate expression of the transgene, animals containing transgenesencoding both the Cre recombinase and a selected protein is required.Such animals can be provided through the construction of“double”transgenic animals, e.g., by mating two transgenic animals, onecontaining a transgene encoding a selected protein and the othercontaining a transgene encoding a recombinase.

[0205] Clones of the non-human transgenic animals described herein canalso be produced according to the methods described in Wilmut, I. et al.Nature 385:810-813(1997) and PCT International Publication Nos. WO97/07668 and WO 97/07669. In brief, a cell, e.g., a somatic cell, fromthe transgenic animal can be isolated and induced to exit the growthcycle and enter G_(o). phase. The quiescent cell can then be fused,e.g., through the use of electrical pulses, to an enucleated oocyte froman animal of the same species from which the quiescent cell is isolated.The reconstructed oocyte is then cultured such that it develops tomorula or blastocyst and then transferred to pseudopregnant femalefoster animal. The offspring born of this female foster animal will be aclone of the animal from which the cell, e.g., the somatic cell, isisolated.

[0206] Transgenic animals containing recombinant cells that express thepeptides described herein are useful to conduct the assays describedherein in an in vivo context. Accordingly, the various physiologicalfactors that are present in vivo and that could effect substratebinding, kinase protein activation, and signal transduction, may not beevident from in vitro cell-free or cell-based assays. Accordingly, it isuseful to provide non-human transgenic animals to assay in vivo kinaseprotein function, including substrate interaction, the effect ofspecific mutant kinase proteins on kinase protein function and substrateinteraction, and the effect of chimeric kinase proteins. It is alsopossible to assess the effect of null mutations, that is, mutations thatsubstantially or completely eliminate one or more kinase proteinfunctions.

[0207] All publications and patents mentioned in the above specificationare herein incorporated by reference. Various modifications andvariations of the described method and system of the invention will beapparent to those skilled in the art without departing from the scopeand spirit of the invention. Although the invention has been describedin connection with specific preferred embodiments, it should beunderstood that the invention as claimed should not be unduly limited tosuch specific embodiments. Indeed, various modifications of theabove-described modes for carrying out the invention which are obviousto those skilled in the field of molecular biology or related fields areintended to be within the scope of the following claims.

1 5 1 9807 DNA Homo sapiens 1 atgcagaaag cccggggcac gcgaggcgaggatgcgggca cgagggcacc ccccagcccc 60 ggagtgcccc cgaaaagggc caaggtgggggccggcggcg gggctcctgt ggccgtggcc 120 ggggcgccag tcttcctgcg gcccctgaagaacgcggcgg tgtgcgcggg cagcgacgtg 180 cggctgcggg tggtggtgag cgggacgccccagcccagcc tccgctggtt ccgggatggg 240 cagctcctgc ccgcgccggc ccccgagcccagctgcctgt ggctgcggcg ctgcggggcg 300 caggacgccg gcgtgtacag ctgcatggcccagaacgagc ggggccgggc ctcctgcgag 360 gcggtgctca cagtgctgga ggtcggagactcagagacgg ctgaggatga catcagcgat 420 gtgcagggaa cccagcgcct ggagcttcgggatgacgggg ccttcagcac ccccacgggg 480 ggttctgaca ccctggtggg cacctccctggacacacccc cgacctccgt gacaggcacc 540 tcagaggagc aagtgagctg gtggggcagcgggcagacgg tcctggagca ggaagcgggc 600 agtgggggtg gcacccgccg cctcccgggcagcccaaggc aagcacaggc aaccggggcc 660 gggccacggc acctgggggt ggagccgctggtgcgggcat ctcgagctaa tctggtgggc 720 gcaagctggg ggtcagagga tagcctttccgtggccagtg acctgtacgg cagcgcattc 780 agcctgtaca gaggacgggc gctctctatccacgtgagcg tccctcagag cgggttgcgc 840 agggaggagc ccgaccttca gcctcaactggccagcgaag ccccacgccg ccctgcccag 900 ccgcctcctt ccaaatccgc gctgctccccccaccgtccc ctcgggtcgg gaagcggtcc 960 ccgccgggac ccccggccca gcccgcggccacccccacgt cgccccaccg tcgcactcag 1020 gagcctgtgc tgcccgagga caccaccaccgaagagaagc gagggaagaa gtccaagtcg 1080 tccgggccct ccctggcggg caccgcggaatcccgacccc agacgccact gagcgaggcc 1140 tcaggccgcc tgtcggcgtt gggccgatcgcctaggctgg tgcgcgccgg ctcccgcatc 1200 ctggacaagc tgcagttctt cgaggagcgacggcgcagcc tggagcgcag cgactcgccg 1260 ccggcgcccc tgcggccctg ggtgcccctgcgcaaggccc gctctctgga gcagcccaag 1320 tcggagcgcg gcgcaccgtg gggcacccccggggcctcgc aggaagaact gcgggcgcca 1380 ggcagcgtgg ccgagcggcg ccgcctgttccagcagaaag cggcctcgct ggacgagcgc 1440 acgcgtcagc gcagcccggc ctcagacctcgagctgcgct tcgcccagga gctgggccgc 1500 atccgccgct ccacgtcgcg ggaggagctggtgcgctcgc acgagtccct gcgcgccacg 1560 ctgcagcgtg ccccatcccc tcgagagcccggcgagcccc cgctcttctc tcggccctcc 1620 acccccaaga catcgcgggc cgtgagccccgccgccgccc agccgccctc tccgagcagc 1680 gcggagaagc cgggggacga gcctgggaggcccaggagcc gcgggccggc gggcaggaca 1740 gagccggggg aaggcccgca gcaggaggttaggcgtcggg accaattccc gctgacccgg 1800 agcagagcca tccaggagtg caggagccctgtgccgcccc ccgccgccga tcccccagag 1860 gccaggacga aagcaccccc cggtcggaagcgggagcccc cggcgcaggc cgtgcgcttc 1920 ctgccctggg ccacgccggg cctggagggcgctgctgtac cccagacctt ggagaagaac 1980 agggcggggc ctgaggcaga gaagaggcttcgcagagggc cggaggagga cggtccctgg 2040 gggccctggg accgccgagg ggcccgcagccagggcaaag gtcgccgggc ccggcccacc 2100 tcccctgagc tcgagtcttc ggatgactcctacgtgtccg ctggagaaga gcccctagag 2160 gcccctgtgt ttgagatccc cctgcagaatgtggtggtgg caccaggggc agatgtgctg 2220 ctcaagtgta tcatcactgc caaccccccgccccaagtgt cctggcacaa ggatgggtca 2280 gcgctgcgca gcgagggccg cctcctcctccgggctgagg gtgagcggca caccctgctg 2340 ctcagggagg ccagggcagc agatgccgggagctatatgg ccaccgccac caacgagctg 2400 ggccaggcca cctgtgccgc ctcactgaccgtgagacccg gtgggtctac atcccctttc 2460 agcagcccca tcacctccga cgaggaatacctgagccccc cagaggagtt cccagagcct 2520 ggggagacct ggccgcgaac ccccaccatgaagcccagtc ccagccagaa ccgccgttct 2580 tctgacactg gctccaaggc accccccaccttcaaggtct cacttatgga ccagtcagta 2640 agagaaggcc aagatgtcat catgagcatccgcgtgcagg gggagcccaa gcctgtggtc 2700 tcctggctga gaaaccgcca gcccgtgcgcccagaccagc ggcgctttgc ggaggaggct 2760 gagggtgggc tgtgccggct gcggatcctggctgcagagc gtggcgatgc tggtttctac 2820 acttgcaaag cggtcaatga gtatggtgctcggcagtgcg aggcccgctt ggaggtccga 2880 gcacaccctg aaagccggtc cctggccgtgctggcccccc tgcaggacgt ggacgtgggg 2940 gccggggaga tggcgctgtt tgagtgcctggtggcggggc ccactgacgt ggaggtggat 3000 tggctgtgcc gtggccgcct gctgcagcctgcactgctca aatgcaagat gcatttcgat 3060 ggccgcaaat gcaagctgct acttacatctgtacatgagg acgacagtgg cgtctacacc 3120 tgcaagctca gcacggccaa agatgagctgacctgcagtg cccggctgac cgtgcggccc 3180 tcgttggcac ccctgttcac acggctgctggaagatgtgg aggtgttgga gggccgagct 3240 gcccgtttcg actgcaagat cagtggcaccccgccccctg ttgttacctg gactcatttt 3300 ggctgcccca tggaggagag tgagaacttgcggctgcggc aggacggggg tctgcactca 3360 ctgcacattg cccatgtggg cagcgaggacgaggggctct atgcggtcag tgctgttaac 3420 acccatggcc aggcccactg ctcagcccagctgtatgtag aagagccccg gacagccgcc 3480 tcaggcccca gctcgaagct ggagaagatgccatccattc ccgaggagcc agagcagggt 3540 gagctggagc ggctgtccat tcccgacttcctgcggccac tgcaggacct ggaggtggga 3600 ctggccaagg aggccatgct agagtgccaggtgaccggcc tgccctaccc caccatcagc 3660 tggttccaca atggccaccg catccagagcagcgacgacc ggcgcatgac acagtacagg 3720 gatgtccatc gcttggtgtt ccctgccgtggggcctcagc acgccggtgt ctacaagagc 3780 gtcattgcca acaagctggg caaagctgcctgctatgccc acctgtatgt cacagatgtg 3840 gtcccaggcc ctccagatgg cgccccgcaggtggtggctg tgacggggag gatggtcaca 3900 ctcacatgga acccccccag gagtctggacatggccatcg acccggactc cctgacgtac 3960 acagtgcagc accaggtgct gggctcggaccagtggacgg cactggtcac aggcctgcgg 4020 gagccagggt gggcagccac agggctgcgtaagggggtcc agcacatctt ccgggtcctc 4080 agcaccactg tcaagagcag cagcaagccctcaccccctt ctgagcctgt gcagctgctg 4140 gagcacggcc caaccctgga ggaggcccctgccatgctgg acaaaccaga catcgtgtat 4200 gtggtggagg gacagcctgc cagcgtcaccgtcacattca accatgtgga ggcccaggtc 4260 gtctggagga gctgccgagg ggccctcctagaggcacggg ccggtgtgta cgagctgagc 4320 cagccagatg atgaccagta ctgtcttcggatctgccggg tgagccgccg ggacatgggg 4380 gccctcacct gcaccgcccg aaaccgtcacggcacacaga cctgctcggt cacattggag 4440 ctggcagagg cccctcggtt tgagtccatcatggaggacg tggaggtggg ggctggggaa 4500 actgctcgct ttgcggtggt ggtcgagggaaaaccactgc cggacatcat gtggtacaag 4560 gacgaggtgc tgctgaccga gagcagccatgtgagcttcg tgtacgagga gaatgagtgc 4620 tccctggtgg tgctcagcac gggggcccaggatggaggcg tctacacctg caccgcccag 4680 aacctggcgg gtgaggtctc ctgcaaagcagagttggctg tgcattcagc tcagacagct 4740 atggaggtcg agggggtcgg ggaggatgaggaccatcgag gaaggagact cagcgacttt 4800 tatgacatcc accaggagat cggcaggggtgctttctcct acttgcggcg catagtggag 4860 cgtagctccg gcctggagtt tgcggccaagttcatcccca gccaggccaa gccaaaggca 4920 tcagcgcgtc gggaggcccg gctgctggccaggctccagc acgactgtgt cctctacttc 4980 catgaggcct tcgagaggcg ccggggactggtcattgtca ccgagctctg cacagaggag 5040 ctgctggagc gaatcgccag gaaacccaccgtgtgtgagt ctgagatccg ggcctatatg 5100 cggcaggtgc tagagggaat acactacctgcaccagagcc acgtgctgca cctcgatgtc 5160 aagcctgaga acctgctggt gtgggatggtgctgcgggcg agcagcaggt gcggatctgt 5220 gactttggga atgcccagga gctgactccaggagagcccc agtactgcca gtatggcaca 5280 cctgagtttg tagcacccga gattgtcaatcagagccccg tgtctggagt cactgacatc 5340 tggcctgtgg gtgttgttgc cttcctctgtctgacaggaa tctccccgtt tgttggggaa 5400 aatgaccgga caacattgat gaacatccgaaactacaacg tggccttcga ggagaccaca 5460 ttcctgagcc tgagcaggga ggcccggggcttcctcatca aagtgttggt gcaggaccgg 5520 ctgagaccta ccgcagaaga gaccctagaacatccttggt tcaaaactca ggcaaagggc 5580 gcagaggtga gcacggatca cctgaagctattcctctccc ggcggaggtg gcagcgctcc 5640 cagatcagct acaaatgcca cctggtgctgcgccccatcc ccgagctgct gcgggccccc 5700 ccagagcggg tgtgggtgac catgcccagaaggccacccc ccagtggggg gctctcatcc 5760 tcctcggatt ctgaagagga agagctggaagagctgccct cagtgccccg cccactgcag 5820 cccgagttct ctggctcccg ggtgtccctcacagacattc ccactgagga tgaggccctg 5880 gggaccccag agactggggc tgccacccccatggactggc aggagcaggg aagggctccc 5940 tctcaggacc aggaggctcc cagcccagaggccctcccct ccccaggcca ggagcccgca 6000 gctggggcta gccccaggcg gggagagctccgcaggggca gctcggctga gagcgccctg 6060 ccccgggccg ggccgcggga gctgggccggggcctgcaca aggcggcgtc tgtggagctg 6120 ccgcagcgcc ggagccccag cccgggagccacccgcctgg cccggggagg cctgggtgag 6180 ggcgagtatg cccagaggct gcaggccctgcgccagcggc tgctgcgggg aggccccgag 6240 gatggcaagg tcagcggcct caggggtcccctgctggaga gcctgggggg ccgtgctcgg 6300 gacccccgga tggcacgagc tgcctccagcgaggcagcgc cccaccacca gcccccactc 6360 gagaaccggg gcctgcaaaa gagcagcagcttctcccagg gtgaggcgga gccccggggc 6420 cggcaccgcc gagcgggggc gcccctcgagatccccgtgg ccaggcttgg ggcccgtagg 6480 ctacaggagt ctccttccct gtctgccctcagcgaggccc agccatccag ccctgcacgg 6540 cccagcgccc ccaaacccag tacccctaagtctgcagaac cttctgccac cacacctagt 6600 gatgctccgc agccccccgc accccagcctgcccaagaca aggctccaga gcccaggcca 6660 gaaccagtcc gagcctccaa gcctgcaccacccccccagg ccctgcaaac cctagcgctg 6720 cccctcacac cctatgctca gatcattcagtccctccagc tgtcaggcca cgcccagggc 6780 ccctcgcagg gccctgccgc gccgccttcagagcccaagc cccacgctgc tgtctttgcc 6840 agggtggcct ccccacctcc gggagcccccgagaagcgcg tgccctcagc cgggggtccc 6900 ccggtgctag ccgagaaagc ccgagttcccacggtgcccc ccaggccagg cagcagtctc 6960 agtagcagca tcgaaaactt ggagtcggaggccgtgttcg aggccaagtt caagcgcagc 7020 cgcgagtcgc ccctgtcgct ggggctgcggctgctgagcc gttcgcgctc ggaggagcgc 7080 ggccccttcc gtggggccga ggaggaggatggcatatacc ggcccagccc ggcggggacc 7140 ccgctggagc tggtgcgacg gcctgagcgctcacgctcgg tgcaggacct cagggctgtc 7200 ggagagcctg gcctcgtccg ccgcctctcgctgtcactgt cccagcggct gcggcggacc 7260 cctcccgcgc agcgccaccc ggcctgggaggcccgcggcg gggacggaga gagctcggag 7320 ggcgggagct cggcgcgggg ctccccggtgctggcgatgc gcaggcggct gagcttcacc 7380 ctggagcggc tgtccagccg attgcagcgcagtggcagca gcgaggactc ggggggcgcg 7440 tcgggccgca gcacgccgct gttcggacggcttcgcaggg ccacgtccga gggcgagagt 7500 ctgcggcgcc ttggccttcc gcacaaccagttggccgccc aggccggcgc caccacgcct 7560 tccgccgagt ccctgggctc cgaggccagcgccacgtcgg gctcctcagc cccaggggaa 7620 agccgaagcc ggctccgctg gggcttctctcggccgcgga aggacaaggg gttatcgcca 7680 ccaaacctct ctgccagcgt ccaggaggagttgggtcacc agtacgtgcg cagtgagtca 7740 gacttccccc cagtcttcca catcaaactcaaggaccagg tgctgctgga gggggaggca 7800 gccaccctgc tctgcctgcc agcggcctgccctgcaccgc acatctcctg gatgaaagac 7860 aagaagtcct tgaggtcaga gccctcagtgatcatcgtgt cctgcaaaga tgggcggcag 7920 ctgctcagca tcccccgggc gggcaagcggcacgccggtc tctatgagtg ctcggccacc 7980 aacgtactgg gcagcatcac cagctcctgtaccgtggctg tggcccgagt cccaggaaag 8040 ctagctcctc cagaggtacc ccagacctaccaggacacgg cgctggtgct gtggaagccg 8100 ggagacagcc gggcaccttg cacgtatacgctggagcggc gagtggatgg ggagtctgtg 8160 tggcaccctg tgagctcagg catccccgactgttactaca acgtgaccca cctgccagtt 8220 ggcgtgactg tgaggttccg tgtggcctgtgccaaccgtg ctgggcaggg gcccttcagc 8280 aactcttctg agaaggtctt tgtcaggggtactcaagatt cttcagctgt gccatctgct 8340 gcccaccaag aggcccctgt cacctcaaggccagccaggg cccggcctcc tgactctcct 8400 acctcactgg ccccacccct agctcctgctgcccccacac ccccgtcagt cactgtcagc 8460 ccctcatctc cccccacacc tcctagccaggccttgtcct cgctcaaggc tgtgggtcca 8520 ccaccccaaa cccctccacg aagacacaggggcctgcagg ctgcccggcc agcggagccc 8580 accctaccca gtacccacgt caccccaagtgagcccaagc ctttcgtcct tgacactggg 8640 accccgatcc cagcctccac tcctcaaggggttaaaccag tgtcttcctc tactcctgtg 8700 tatgtggtga cttcctttgt gtctgcaccaccagcccctg agcccccagc ccctgagccc 8760 cctcctgagc ctaccaaggt gactgtgcagagcctcagcc cggccaagga ggtggtcagc 8820 tcccctggga gcagtccccg aagctctcccaggcctgagg gtaccactct tcgacagggt 8880 ccccctcaga aaccctacac cttcctggaggagaaagcca ggcagggccg ctttggtgtt 8940 gtgcgagcgt gccgggagaa tgccacggggcgaacgttcg tggccaagat cgtgccctat 9000 gctgccgagg gcaagcggcg ggtcctgcaggagtacgagg tgctgcggac cctgcaccac 9060 gagcggatca tgtccctgca cgaggcctacatcacccctc ggtacctcgt gctcattgct 9120 gagagctgtg gcaaccggga actcctctgtgggctcagtg acaggttccg gtattctgag 9180 gatgacgtgg ccacttacat ggtgcagctgctacaaggcc tggactacct ccacggccac 9240 cacgtgctcc acctagacat caagccagacaacctgctgc tggcccctga caatgccctc 9300 aagattgtgg actttggcag tgcccagccctacaaccccc aggcccttag gccccttggc 9360 caccgcacgg gcacgctgga gttcatggctccggagatgg tgaagggaga acccatcggc 9420 tctgccacgg acatctgggg agcgggtgtgctcacttaca ttatgctcag tggacgctcc 9480 ccgttctatg agccagaccc ccaggaaacggaggctcgga ttgtgggggg ccgctttgat 9540 gccttccagc tgtaccccaa tacatcccagagcgccaccc tcttcttgcg aaaggttctc 9600 tctgtacatc cctggagccg gccctccctgcaggactgcc tggcccaccc atggttgcag 9660 gacgcctacc tgatgaagct gcgccgccagacgctcacct tcaccaccaa ccggctcaag 9720 gagttcctgg gcgagcagcg gcggcgccgggctgaggctg ccacccgcca caaggtgctg 9780 ctgcgctcct accctggcgg cccctag 98072 3268 PRT Homo sapiens 2 Met Gln Lys Ala Arg Gly Thr Arg Gly Glu AspAla Gly Thr Arg Ala 1 5 10 15 Pro Pro Ser Pro Gly Val Pro Pro Lys ArgAla Lys Val Gly Ala Gly 20 25 30 Gly Gly Ala Pro Val Ala Val Ala Gly AlaPro Val Phe Leu Arg Pro 35 40 45 Leu Lys Asn Ala Ala Val Cys Ala Gly SerAsp Val Arg Leu Arg Val 50 55 60 Val Val Ser Gly Thr Pro Gln Pro Ser LeuArg Trp Phe Arg Asp Gly 65 70 75 80 Gln Leu Leu Pro Ala Pro Ala Pro GluPro Ser Cys Leu Trp Leu Arg 85 90 95 Arg Cys Gly Ala Gln Asp Ala Gly ValTyr Ser Cys Met Ala Gln Asn 100 105 110 Glu Arg Gly Arg Ala Ser Cys GluAla Val Leu Thr Val Leu Glu Val 115 120 125 Gly Asp Ser Glu Thr Ala GluAsp Asp Ile Ser Asp Val Gln Gly Thr 130 135 140 Gln Arg Leu Glu Leu ArgAsp Asp Gly Ala Phe Ser Thr Pro Thr Gly 145 150 155 160 Gly Ser Asp ThrLeu Val Gly Thr Ser Leu Asp Thr Pro Pro Thr Ser 165 170 175 Val Thr GlyThr Ser Glu Glu Gln Val Ser Trp Trp Gly Ser Gly Gln 180 185 190 Thr ValLeu Glu Gln Glu Ala Gly Ser Gly Gly Gly Thr Arg Arg Leu 195 200 205 ProGly Ser Pro Arg Gln Ala Gln Ala Thr Gly Ala Gly Pro Arg His 210 215 220Leu Gly Val Glu Pro Leu Val Arg Ala Ser Arg Ala Asn Leu Val Gly 225 230235 240 Ala Ser Trp Gly Ser Glu Asp Ser Leu Ser Val Ala Ser Asp Leu Tyr245 250 255 Gly Ser Ala Phe Ser Leu Tyr Arg Gly Arg Ala Leu Ser Ile HisVal 260 265 270 Ser Val Pro Gln Ser Gly Leu Arg Arg Glu Glu Pro Asp LeuGln Pro 275 280 285 Gln Leu Ala Ser Glu Ala Pro Arg Arg Pro Ala Gln ProPro Pro Ser 290 295 300 Lys Ser Ala Leu Leu Pro Pro Pro Ser Pro Arg ValGly Lys Arg Ser 305 310 315 320 Pro Pro Gly Pro Pro Ala Gln Pro Ala AlaThr Pro Thr Ser Pro His 325 330 335 Arg Arg Thr Gln Glu Pro Val Leu ProGlu Asp Thr Thr Thr Glu Glu 340 345 350 Lys Arg Gly Lys Lys Ser Lys SerSer Gly Pro Ser Leu Ala Gly Thr 355 360 365 Ala Glu Ser Arg Pro Gln ThrPro Leu Ser Glu Ala Ser Gly Arg Leu 370 375 380 Ser Ala Leu Gly Arg SerPro Arg Leu Val Arg Ala Gly Ser Arg Ile 385 390 395 400 Leu Asp Lys LeuGln Phe Phe Glu Glu Arg Arg Arg Ser Leu Glu Arg 405 410 415 Ser Asp SerPro Pro Ala Pro Leu Arg Pro Trp Val Pro Leu Arg Lys 420 425 430 Ala ArgSer Leu Glu Gln Pro Lys Ser Glu Arg Gly Ala Pro Trp Gly 435 440 445 ThrPro Gly Ala Ser Gln Glu Glu Leu Arg Ala Pro Gly Ser Val Ala 450 455 460Glu Arg Arg Arg Leu Phe Gln Gln Lys Ala Ala Ser Leu Asp Glu Arg 465 470475 480 Thr Arg Gln Arg Ser Pro Ala Ser Asp Leu Glu Leu Arg Phe Ala Gln485 490 495 Glu Leu Gly Arg Ile Arg Arg Ser Thr Ser Arg Glu Glu Leu ValArg 500 505 510 Ser His Glu Ser Leu Arg Ala Thr Leu Gln Arg Ala Pro SerPro Arg 515 520 525 Glu Pro Gly Glu Pro Pro Leu Phe Ser Arg Pro Ser ThrPro Lys Thr 530 535 540 Ser Arg Ala Val Ser Pro Ala Ala Ala Gln Pro ProSer Pro Ser Ser 545 550 555 560 Ala Glu Lys Pro Gly Asp Glu Pro Gly ArgPro Arg Ser Arg Gly Pro 565 570 575 Ala Gly Arg Thr Glu Pro Gly Glu GlyPro Gln Gln Glu Val Arg Arg 580 585 590 Arg Asp Gln Phe Pro Leu Thr ArgSer Arg Ala Ile Gln Glu Cys Arg 595 600 605 Ser Pro Val Pro Pro Pro AlaAla Asp Pro Pro Glu Ala Arg Thr Lys 610 615 620 Ala Pro Pro Gly Arg LysArg Glu Pro Pro Ala Gln Ala Val Arg Phe 625 630 635 640 Leu Pro Trp AlaThr Pro Gly Leu Glu Gly Ala Ala Val Pro Gln Thr 645 650 655 Leu Glu LysAsn Arg Ala Gly Pro Glu Ala Glu Lys Arg Leu Arg Arg 660 665 670 Gly ProGlu Glu Asp Gly Pro Trp Gly Pro Trp Asp Arg Arg Gly Ala 675 680 685 ArgSer Gln Gly Lys Gly Arg Arg Ala Arg Pro Thr Ser Pro Glu Leu 690 695 700Glu Ser Ser Asp Asp Ser Tyr Val Ser Ala Gly Glu Glu Pro Leu Glu 705 710715 720 Ala Pro Val Phe Glu Ile Pro Leu Gln Asn Val Val Val Ala Pro Gly725 730 735 Ala Asp Val Leu Leu Lys Cys Ile Ile Thr Ala Asn Pro Pro ProGln 740 745 750 Val Ser Trp His Lys Asp Gly Ser Ala Leu Arg Ser Glu GlyArg Leu 755 760 765 Leu Leu Arg Ala Glu Gly Glu Arg His Thr Leu Leu LeuArg Glu Ala 770 775 780 Arg Ala Ala Asp Ala Gly Ser Tyr Met Ala Thr AlaThr Asn Glu Leu 785 790 795 800 Gly Gln Ala Thr Cys Ala Ala Ser Leu ThrVal Arg Pro Gly Gly Ser 805 810 815 Thr Ser Pro Phe Ser Ser Pro Ile ThrSer Asp Glu Glu Tyr Leu Ser 820 825 830 Pro Pro Glu Glu Phe Pro Glu ProGly Glu Thr Trp Pro Arg Thr Pro 835 840 845 Thr Met Lys Pro Ser Pro SerGln Asn Arg Arg Ser Ser Asp Thr Gly 850 855 860 Ser Lys Ala Pro Pro ThrPhe Lys Val Ser Leu Met Asp Gln Ser Val 865 870 875 880 Arg Glu Gly GlnAsp Val Ile Met Ser Ile Arg Val Gln Gly Glu Pro 885 890 895 Lys Pro ValVal Ser Trp Leu Arg Asn Arg Gln Pro Val Arg Pro Asp 900 905 910 Gln ArgArg Phe Ala Glu Glu Ala Glu Gly Gly Leu Cys Arg Leu Arg 915 920 925 IleLeu Ala Ala Glu Arg Gly Asp Ala Gly Phe Tyr Thr Cys Lys Ala 930 935 940Val Asn Glu Tyr Gly Ala Arg Gln Cys Glu Ala Arg Leu Glu Val Arg 945 950955 960 Ala His Pro Glu Ser Arg Ser Leu Ala Val Leu Ala Pro Leu Gln Asp965 970 975 Val Asp Val Gly Ala Gly Glu Met Ala Leu Phe Glu Cys Leu ValAla 980 985 990 Gly Pro Thr Asp Val Glu Val Asp Trp Leu Cys Arg Gly ArgLeu Leu 995 1000 1005 Gln Pro Ala Leu Leu Lys Cys Lys Met His Phe AspGly Arg Lys Cys 1010 1015 1020 Lys Leu Leu Leu Thr Ser Val His Glu AspAsp Ser Gly Val Tyr Thr 1025 1030 1035 1040 Cys Lys Leu Ser Thr Ala LysAsp Glu Leu Thr Cys Ser Ala Arg Leu 1045 1050 1055 Thr Val Arg Pro SerLeu Ala Pro Leu Phe Thr Arg Leu Leu Glu Asp 1060 1065 1070 Val Glu ValLeu Glu Gly Arg Ala Ala Arg Phe Asp Cys Lys Ile Ser 1075 1080 1085 GlyThr Pro Pro Pro Val Val Thr Trp Thr His Phe Gly Cys Pro Met 1090 10951100 Glu Glu Ser Glu Asn Leu Arg Leu Arg Gln Asp Gly Gly Leu His Ser1105 1110 1115 1120 Leu His Ile Ala His Val Gly Ser Glu Asp Glu Gly LeuTyr Ala Val 1125 1130 1135 Ser Ala Val Asn Thr His Gly Gln Ala His CysSer Ala Gln Leu Tyr 1140 1145 1150 Val Glu Glu Pro Arg Thr Ala Ala SerGly Pro Ser Ser Lys Leu Glu 1155 1160 1165 Lys Met Pro Ser Ile Pro GluGlu Pro Glu Gln Gly Glu Leu Glu Arg 1170 1175 1180 Leu Ser Ile Pro AspPhe Leu Arg Pro Leu Gln Asp Leu Glu Val Gly 1185 1190 1195 1200 Leu AlaLys Glu Ala Met Leu Glu Cys Gln Val Thr Gly Leu Pro Tyr 1205 1210 1215Pro Thr Ile Ser Trp Phe His Asn Gly His Arg Ile Gln Ser Ser Asp 12201225 1230 Asp Arg Arg Met Thr Gln Tyr Arg Asp Val His Arg Leu Val PhePro 1235 1240 1245 Ala Val Gly Pro Gln His Ala Gly Val Tyr Lys Ser ValIle Ala Asn 1250 1255 1260 Lys Leu Gly Lys Ala Ala Cys Tyr Ala His LeuTyr Val Thr Asp Val 1265 1270 1275 1280 Val Pro Gly Pro Pro Asp Gly AlaPro Gln Val Val Ala Val Thr Gly 1285 1290 1295 Arg Met Val Thr Leu ThrTrp Asn Pro Pro Arg Ser Leu Asp Met Ala 1300 1305 1310 Ile Asp Pro AspSer Leu Thr Tyr Thr Val Gln His Gln Val Leu Gly 1315 1320 1325 Ser AspGln Trp Thr Ala Leu Val Thr Gly Leu Arg Glu Pro Gly Trp 1330 1335 1340Ala Ala Thr Gly Leu Arg Lys Gly Val Gln His Ile Phe Arg Val Leu 13451350 1355 1360 Ser Thr Thr Val Lys Ser Ser Ser Lys Pro Ser Pro Pro SerGlu Pro 1365 1370 1375 Val Gln Leu Leu Glu His Gly Pro Thr Leu Glu GluAla Pro Ala Met 1380 1385 1390 Leu Asp Lys Pro Asp Ile Val Tyr Val ValGlu Gly Gln Pro Ala Ser 1395 1400 1405 Val Thr Val Thr Phe Asn His ValGlu Ala Gln Val Val Trp Arg Ser 1410 1415 1420 Cys Arg Gly Ala Leu LeuGlu Ala Arg Ala Gly Val Tyr Glu Leu Ser 1425 1430 1435 1440 Gln Pro AspAsp Asp Gln Tyr Cys Leu Arg Ile Cys Arg Val Ser Arg 1445 1450 1455 ArgAsp Met Gly Ala Leu Thr Cys Thr Ala Arg Asn Arg His Gly Thr 1460 14651470 Gln Thr Cys Ser Val Thr Leu Glu Leu Ala Glu Ala Pro Arg Phe Glu1475 1480 1485 Ser Ile Met Glu Asp Val Glu Val Gly Ala Gly Glu Thr AlaArg Phe 1490 1495 1500 Ala Val Val Val Glu Gly Lys Pro Leu Pro Asp IleMet Trp Tyr Lys 1505 1510 1515 1520 Asp Glu Val Leu Leu Thr Glu Ser SerHis Val Ser Phe Val Tyr Glu 1525 1530 1535 Glu Asn Glu Cys Ser Leu ValVal Leu Ser Thr Gly Ala Gln Asp Gly 1540 1545 1550 Gly Val Tyr Thr CysThr Ala Gln Asn Leu Ala Gly Glu Val Ser Cys 1555 1560 1565 Lys Ala GluLeu Ala Val His Ser Ala Gln Thr Ala Met Glu Val Glu 1570 1575 1580 GlyVal Gly Glu Asp Glu Asp His Arg Gly Arg Arg Leu Ser Asp Phe 1585 15901595 1600 Tyr Asp Ile His Gln Glu Ile Gly Arg Gly Ala Phe Ser Tyr LeuArg 1605 1610 1615 Arg Ile Val Glu Arg Ser Ser Gly Leu Glu Phe Ala AlaLys Phe Ile 1620 1625 1630 Pro Ser Gln Ala Lys Pro Lys Ala Ser Ala ArgArg Glu Ala Arg Leu 1635 1640 1645 Leu Ala Arg Leu Gln His Asp Cys ValLeu Tyr Phe His Glu Ala Phe 1650 1655 1660 Glu Arg Arg Arg Gly Leu ValIle Val Thr Glu Leu Cys Thr Glu Glu 1665 1670 1675 1680 Leu Leu Glu ArgIle Ala Arg Lys Pro Thr Val Cys Glu Ser Glu Ile 1685 1690 1695 Arg AlaTyr Met Arg Gln Val Leu Glu Gly Ile His Tyr Leu His Gln 1700 1705 1710Ser His Val Leu His Leu Asp Val Lys Pro Glu Asn Leu Leu Val Trp 17151720 1725 Asp Gly Ala Ala Gly Glu Gln Gln Val Arg Ile Cys Asp Phe GlyAsn 1730 1735 1740 Ala Gln Glu Leu Thr Pro Gly Glu Pro Gln Tyr Cys GlnTyr Gly Thr 1745 1750 1755 1760 Pro Glu Phe Val Ala Pro Glu Ile Val AsnGln Ser Pro Val Ser Gly 1765 1770 1775 Val Thr Asp Ile Trp Pro Val GlyVal Val Ala Phe Leu Cys Leu Thr 1780 1785 1790 Gly Ile Ser Pro Phe ValGly Glu Asn Asp Arg Thr Thr Leu Met Asn 1795 1800 1805 Ile Arg Asn TyrAsn Val Ala Phe Glu Glu Thr Thr Phe Leu Ser Leu 1810 1815 1820 Ser ArgGlu Ala Arg Gly Phe Leu Ile Lys Val Leu Val Gln Asp Arg 1825 1830 18351840 Leu Arg Pro Thr Ala Glu Glu Thr Leu Glu His Pro Trp Phe Lys Thr1845 1850 1855 Gln Ala Lys Gly Ala Glu Val Ser Thr Asp His Leu Lys LeuPhe Leu 1860 1865 1870 Ser Arg Arg Arg Trp Gln Arg Ser Gln Ile Ser TyrLys Cys His Leu 1875 1880 1885 Val Leu Arg Pro Ile Pro Glu Leu Leu ArgAla Pro Pro Glu Arg Val 1890 1895 1900 Trp Val Thr Met Pro Arg Arg ProPro Pro Ser Gly Gly Leu Ser Ser 1905 1910 1915 1920 Ser Ser Asp Ser GluGlu Glu Glu Leu Glu Glu Leu Pro Ser Val Pro 1925 1930 1935 Arg Pro LeuGln Pro Glu Phe Ser Gly Ser Arg Val Ser Leu Thr Asp 1940 1945 1950 IlePro Thr Glu Asp Glu Ala Leu Gly Thr Pro Glu Thr Gly Ala Ala 1955 19601965 Thr Pro Met Asp Trp Gln Glu Gln Gly Arg Ala Pro Ser Gln Asp Gln1970 1975 1980 Glu Ala Pro Ser Pro Glu Ala Leu Pro Ser Pro Gly Gln GluPro Ala 1985 1990 1995 2000 Ala Gly Ala Ser Pro Arg Arg Gly Glu Leu ArgArg Gly Ser Ser Ala 2005 2010 2015 Glu Ser Ala Leu Pro Arg Ala Gly ProArg Glu Leu Gly Arg Gly Leu 2020 2025 2030 His Lys Ala Ala Ser Val GluLeu Pro Gln Arg Arg Ser Pro Ser Pro 2035 2040 2045 Gly Ala Thr Arg LeuAla Arg Gly Gly Leu Gly Glu Gly Glu Tyr Ala 2050 2055 2060 Gln Arg LeuGln Ala Leu Arg Gln Arg Leu Leu Arg Gly Gly Pro Glu 2065 2070 2075 2080Asp Gly Lys Val Ser Gly Leu Arg Gly Pro Leu Leu Glu Ser Leu Gly 20852090 2095 Gly Arg Ala Arg Asp Pro Arg Met Ala Arg Ala Ala Ser Ser GluAla 2100 2105 2110 Ala Pro His His Gln Pro Pro Leu Glu Asn Arg Gly LeuGln Lys Ser 2115 2120 2125 Ser Ser Phe Ser Gln Gly Glu Ala Glu Pro ArgGly Arg His Arg Arg 2130 2135 2140 Ala Gly Ala Pro Leu Glu Ile Pro ValAla Arg Leu Gly Ala Arg Arg 2145 2150 2155 2160 Leu Gln Glu Ser Pro SerLeu Ser Ala Leu Ser Glu Ala Gln Pro Ser 2165 2170 2175 Ser Pro Ala ArgPro Ser Ala Pro Lys Pro Ser Thr Pro Lys Ser Ala 2180 2185 2190 Glu ProSer Ala Thr Thr Pro Ser Asp Ala Pro Gln Pro Pro Ala Pro 2195 2200 2205Gln Pro Ala Gln Asp Lys Ala Pro Glu Pro Arg Pro Glu Pro Val Arg 22102215 2220 Ala Ser Lys Pro Ala Pro Pro Pro Gln Ala Leu Gln Thr Leu AlaLeu 2225 2230 2235 2240 Pro Leu Thr Pro Tyr Ala Gln Ile Ile Gln Ser LeuGln Leu Ser Gly 2245 2250 2255 His Ala Gln Gly Pro Ser Gln Gly Pro AlaAla Pro Pro Ser Glu Pro 2260 2265 2270 Lys Pro His Ala Ala Val Phe AlaArg Val Ala Ser Pro Pro Pro Gly 2275 2280 2285 Ala Pro Glu Lys Arg ValPro Ser Ala Gly Gly Pro Pro Val Leu Ala 2290 2295 2300 Glu Lys Ala ArgVal Pro Thr Val Pro Pro Arg Pro Gly Ser Ser Leu 2305 2310 2315 2320 SerSer Ser Ile Glu Asn Leu Glu Ser Glu Ala Val Phe Glu Ala Lys 2325 23302335 Phe Lys Arg Ser Arg Glu Ser Pro Leu Ser Leu Gly Leu Arg Leu Leu2340 2345 2350 Ser Arg Ser Arg Ser Glu Glu Arg Gly Pro Phe Arg Gly AlaGlu Glu 2355 2360 2365 Glu Asp Gly Ile Tyr Arg Pro Ser Pro Ala Gly ThrPro Leu Glu Leu 2370 2375 2380 Val Arg Arg Pro Glu Arg Ser Arg Ser ValGln Asp Leu Arg Ala Val 2385 2390 2395 2400 Gly Glu Pro Gly Leu Val ArgArg Leu Ser Leu Ser Leu Ser Gln Arg 2405 2410 2415 Leu Arg Arg Thr ProPro Ala Gln Arg His Pro Ala Trp Glu Ala Arg 2420 2425 2430 Gly Gly AspGly Glu Ser Ser Glu Gly Gly Ser Ser Ala Arg Gly Ser 2435 2440 2445 ProVal Leu Ala Met Arg Arg Arg Leu Ser Phe Thr Leu Glu Arg Leu 2450 24552460 Ser Ser Arg Leu Gln Arg Ser Gly Ser Ser Glu Asp Ser Gly Gly Ala2465 2470 2475 2480 Ser Gly Arg Ser Thr Pro Leu Phe Gly Arg Leu Arg ArgAla Thr Ser 2485 2490 2495 Glu Gly Glu Ser Leu Arg Arg Leu Gly Leu ProHis Asn Gln Leu Ala 2500 2505 2510 Ala Gln Ala Gly Ala Thr Thr Pro SerAla Glu Ser Leu Gly Ser Glu 2515 2520 2525 Ala Ser Ala Thr Ser Gly SerSer Ala Pro Gly Glu Ser Arg Ser Arg 2530 2535 2540 Leu Arg Trp Gly PheSer Arg Pro Arg Lys Asp Lys Gly Leu Ser Pro 2545 2550 2555 2560 Pro AsnLeu Ser Ala Ser Val Gln Glu Glu Leu Gly His Gln Tyr Val 2565 2570 2575Arg Ser Glu Ser Asp Phe Pro Pro Val Phe His Ile Lys Leu Lys Asp 25802585 2590 Gln Val Leu Leu Glu Gly Glu Ala Ala Thr Leu Leu Cys Leu ProAla 2595 2600 2605 Ala Cys Pro Ala Pro His Ile Ser Trp Met Lys Asp LysLys Ser Leu 2610 2615 2620 Arg Ser Glu Pro Ser Val Ile Ile Val Ser CysLys Asp Gly Arg Gln 2625 2630 2635 2640 Leu Leu Ser Ile Pro Arg Ala GlyLys Arg His Ala Gly Leu Tyr Glu 2645 2650 2655 Cys Ser Ala Thr Asn ValLeu Gly Ser Ile Thr Ser Ser Cys Thr Val 2660 2665 2670 Ala Val Ala ArgVal Pro Gly Lys Leu Ala Pro Pro Glu Val Pro Gln 2675 2680 2685 Thr TyrGln Asp Thr Ala Leu Val Leu Trp Lys Pro Gly Asp Ser Arg 2690 2695 2700Ala Pro Cys Thr Tyr Thr Leu Glu Arg Arg Val Asp Gly Glu Ser Val 27052710 2715 2720 Trp His Pro Val Ser Ser Gly Ile Pro Asp Cys Tyr Tyr AsnVal Thr 2725 2730 2735 His Leu Pro Val Gly Val Thr Val Arg Phe Arg ValAla Cys Ala Asn 2740 2745 2750 Arg Ala Gly Gln Gly Pro Phe Ser Asn SerSer Glu Lys Val Phe Val 2755 2760 2765 Arg Gly Thr Gln Asp Ser Ser AlaVal Pro Ser Ala Ala His Gln Glu 2770 2775 2780 Ala Pro Val Thr Ser ArgPro Ala Arg Ala Arg Pro Pro Asp Ser Pro 2785 2790 2795 2800 Thr Ser LeuAla Pro Pro Leu Ala Pro Ala Ala Pro Thr Pro Pro Ser 2805 2810 2815 ValThr Val Ser Pro Ser Ser Pro Pro Thr Pro Pro Ser Gln Ala Leu 2820 28252830 Ser Ser Leu Lys Ala Val Gly Pro Pro Pro Gln Thr Pro Pro Arg Arg2835 2840 2845 His Arg Gly Leu Gln Ala Ala Arg Pro Ala Glu Pro Thr LeuPro Ser 2850 2855 2860 Thr His Val Thr Pro Ser Glu Pro Lys Pro Phe ValLeu Asp Thr Gly 2865 2870 2875 2880 Thr Pro Ile Pro Ala Ser Thr Pro GlnGly Val Lys Pro Val Ser Ser 2885 2890 2895 Ser Thr Pro Val Tyr Val ValThr Ser Phe Val Ser Ala Pro Pro Ala 2900 2905 2910 Pro Glu Pro Pro AlaPro Glu Pro Pro Pro Glu Pro Thr Lys Val Thr 2915 2920 2925 Val Gln SerLeu Ser Pro Ala Lys Glu Val Val Ser Ser Pro Gly Ser 2930 2935 2940 SerPro Arg Ser Ser Pro Arg Pro Glu Gly Thr Thr Leu Arg Gln Gly 2945 29502955 2960 Pro Pro Gln Lys Pro Tyr Thr Phe Leu Glu Glu Lys Ala Arg GlnGly 2965 2970 2975 Arg Phe Gly Val Val Arg Ala Cys Arg Glu Asn Ala ThrGly Arg Thr 2980 2985 2990 Phe Val Ala Lys Ile Val Pro Tyr Ala Ala GluGly Lys Arg Arg Val 2995 3000 3005 Leu Gln Glu Tyr Glu Val Leu Arg ThrLeu His His Glu Arg Ile Met 3010 3015 3020 Ser Leu His Glu Ala Tyr IleThr Pro Arg Tyr Leu Val Leu Ile Ala 3025 3030 3035 3040 Glu Ser Cys GlyAsn Arg Glu Leu Leu Cys Gly Leu Ser Asp Arg Phe 3045 3050 3055 Arg TyrSer Glu Asp Asp Val Ala Thr Tyr Met Val Gln Leu Leu Gln 3060 3065 3070Gly Leu Asp Tyr Leu His Gly His His Val Leu His Leu Asp Ile Lys 30753080 3085 Pro Asp Asn Leu Leu Leu Ala Pro Asp Asn Ala Leu Lys Ile ValAsp 3090 3095 3100 Phe Gly Ser Ala Gln Pro Tyr Asn Pro Gln Ala Leu ArgPro Leu Gly 3105 3110 3115 3120 His Arg Thr Gly Thr Leu Glu Phe Met AlaPro Glu Met Val Lys Gly 3125 3130 3135 Glu Pro Ile Gly Ser Ala Thr AspIle Trp Gly Ala Gly Val Leu Thr 3140 3145 3150 Tyr Ile Met Leu Ser GlyArg Ser Pro Phe Tyr Glu Pro Asp Pro Gln 3155 3160 3165 Glu Thr Glu AlaArg Ile Val Gly Gly Arg Phe Asp Ala Phe Gln Leu 3170 3175 3180 Tyr ProAsn Thr Ser Gln Ser Ala Thr Leu Phe Leu Arg Lys Val Leu 3185 3190 31953200 Ser Val His Pro Trp Ser Arg Pro Ser Leu Gln Asp Cys Leu Ala His3205 3210 3215 Pro Trp Leu Gln Asp Ala Tyr Leu Met Lys Leu Arg Arg GlnThr Leu 3220 3225 3230 Thr Phe Thr Thr Asn Arg Leu Lys Glu Phe Leu GlyGlu Gln Arg Arg 3235 3240 3245 Arg Arg Ala Glu Ala Ala Thr Arg His LysVal Leu Leu Arg Ser Tyr 3250 3255 3260 Pro Gly Gly Pro 3265 3 62805 DNAHomo sapiens 3 ctttgtctgt tcactgctat atccctagtc cctagcacag tgccagtacatagtagaaac 60 tcaaaaatat ttgtggatga ataaataaaa aaattatgga tgaataaataattaaaaccc 120 tgagttgtgc tacctccatt ctatagatga ggaaaccgag gcttagagatgctaggtaac 180 ttgcttgaga tcgcatcgtt catttattca accaacttac taaccagccaacatttacag 240 tctacccact gcattccaca cacatttaga ggcacagtgt tgggtggctttgggttattt 300 gtttttcgaa atactctcta ttcctttttt cttgatatac catctgtttgcaatgacttc 360 ccccatattg tcaccttcta gaattcaatt tacactttag aattcgattcatcatcagtg 420 gttgccagag gttggtggag ggggtctatg gtagagtcta tgactccgaagggggtacat 480 gagctagtat ttggggtgat ggaattgttt gctctgtatg gtcctggagtggcagataca 540 tgattctatg catttgtcaa aacccagaga actgcacctc ataaaaaaataaactttagg 600 ccaggcacgg tggctcacac ctgtaatccc agtgctttag gaggctgaggcaggcagatc 660 acctgaggtc gggagttcga gaccagcccg accaaaatgg agaaaccccgtctctactaa 720 aaatacaaaa ttagctgggt gtggtggtgc atgcctgtaa tcccagctactcgggaggct 780 gaggcaggag aattgctcga acccgggagg aggaggttgt ggtgagccgagatcacacca 840 ttgcactccg gcctgggcaa caagagtgaa actccgtctc aaaaaaaaaaaaaaacttta 900 atgtatgcaa actgtaaaaa aaattattcc tcaaggttct taacctctatggatgtaatt 960 cagtgtttaa atggttcctc ctataccttt tacacactgt ctctcgcgctctctctcttt 1020 ctctttgact tcagtatccc agaatgagga tggggaagag gaggcaagggtaagagtaac 1080 attctctgcc tctgaatact catggctcct ctcagccctt cctgggtttcatccctcagg 1140 gctcaaggtc aggcctgggt ctcctacttg gacttcttaa aaaattttttactttatgat 1200 aactgtagat tcacaggcaa ttataagaaa taatgcagag agatcctgaattaccttcac 1260 ttggtttcct cctagggtaa catcttgtat gactatagta cagcatcacaaccaggaaag 1320 gggcattggt ataatccacc taccttctgc aagttttacc agtgttacatgtactgttag 1380 tgttgcgtac aagtgcaaat gcacatttag ttctatgcaa ctttatcacttgtgtagatc 1440 cacataacca ccaccattac cactgtcaag atacagaact gttctgtttttgttttgttt 1500 tgttttttga gacagagtct cactctgttg cccaggctgc agtgcagtggtgccatctag 1560 gctcactgca acctccacct cccaggttca agcgattctc ctgcctcagcctcctgagta 1620 gctgggacta caggcatgag ccactacacc tggctaattt tttgtatttttagtagagac 1680 aggtttcacc atgttggcca ggctggtgtt gaattcctga gttcaagtgatccacccacc 1740 tcggcctccc aaagtgctgg aattacaggt gttagccacc gcgcccagccaagatacaga 1800 actgttctat cacaaaggtc tcatctggac tcttgatgtt tctcaacgtgcaacacttag 1860 gcacatcaga atcagttgag tcatttgtta aatgtgcaga ttcctcccagctcagctgct 1920 gaaccagtgt ggggcaagga ggctgggaat ctggccttta cttgaactggccgtcatttc 1980 tatccatcct ccactttgtg gccgccaaga ggatcctcct aagacacagctcccaccgtg 2040 ttttttctgc cgcttaaagc tgtgtagtgg cgccccctgc tttcagggtagagaaaccaa 2100 agccttagca aacaaagcct tctccaggcc ccacctccct tcttccactcaccccaccca 2160 ctacgcttca atccctccaa acctctctaa ttcccaggac gcctttgttttcatcaggct 2220 cattttgttc atgccgttcc ctctgcctgg aatgtcctgc ccactcttttctgcctattg 2280 caaccctatt ccaccaccca ttcttacaga tgaggacttg gaggttgagagaggtttggt 2340 gtcacccagc aagtaagggc agggcgcagt ggaggcccca atccacctgactcccaggct 2400 cctggtctta ctgttcgcca gctgtaatgg aggcgctggg ggaggccatggtccctcttc 2460 ggagctgtct gctcacctcc acttgggcct gcctccccat ccacctctcaggcatctcac 2520 caggaccgtt cctcttcttc ccctcccagc gaagccgggc agggatgagggttctgagat 2580 gagggaggaa gggaaatggg attgagccca ggggtaacct gactccctgcagtgggtcgt 2640 gtgggggcca ggcacactac ggaggggaaa gcctggaaca aataccgagggactccctta 2700 agccgggccg gcgatggggg ctcctggagg gagagaagga gccaagtggagtcaagtccc 2760 tcccctgctg ccccctccct ccacggctcc ctcgcaaccc gagccggggggcctaaaaat 2820 agcccccagg cgcaatcgcc tgccgccccg gtgaccttct gggtagcacaggccgaaggc 2880 gggcgggcag caggaaggca ggccgccggc cccccagact tgtctcctagggcaccgtcc 2940 cgcgggtgcc cccgtggccg cccagttccg gcgtcccccc agcccagctctcagtggcca 3000 tgcagaaagc ccggggcacg cgaggcgagg atgcgggcac gagggcaccccccagccccg 3060 gagtgccccc gaaaagggcc aaggtggggg ccggcggcgg ggctcctgtggccgtggccg 3120 gggcgccagt cttcctgcgg cccctgaaga acgcggcggt gtgcgcgggcagcgacgtgc 3180 ggctgcgggt ggtggtgagc gggacgcccc agcccagcct ccgctggttccgggatgggc 3240 agctcctgcc cgcgccggcc cccgagccca gctgcctgtg gctgcggcgctgcggggcgc 3300 aggacgccgg cgtgtacagc tgcatggccc agaacgagcg gggccgggcctcctgcgagg 3360 cggtgctcac agtgctggag gtcggaggta aagggcaggt gggggccgcgcccggcaggg 3420 gcggggtgct cagaggtaga aaagggctgc ccaggccacg cgggtaaggtactggatact 3480 ggttccgccg ccttcttccc aggtgccctg gcttctcggc tgcccggccccagaagtgag 3540 acgaagagcc aagtgcaggg aatggggtgt caaggtagag aggctccccacaggaaggtc 3600 agaggtcaag gggcagcaag cggttgataa gccaagcctg agacccactcccacctctca 3660 gggaattctg gggtggaagt tcttctcctc ctgtggagaa aagcctcctgggggaaaggg 3720 tgtccttcag ttccatgatt taaacttgag attgacactc cgatcagctccttaacaggg 3780 gagtccatgt ccgaaggagg gggccagctc ctctggtcca ggctgcactgttgaagggat 3840 ggctcagagc tccctgcagg ccattgccgt ggcagggttg atgtggtcagctctaggtgg 3900 ggtgtggaag aggcctatgg ttggccacgt gtgaacaggg tccagggtaggaggaggtgg 3960 agcccgagcc agggccccat gggcatgaat ggaggtgagt gcttgagaatccacatgcag 4020 gtgtgtgcct gcatgggtgc tgtggagggc cctggattct gtgtggtggtgcaaacaggt 4080 gaggtatggg cacgtggagc tggaatggga agctcctgga ccatgtctacctgagcttcc 4140 agagtggatg tttccagagc atggaagggg gatgctatgg accagtgcttgtcccccgcc 4200 catagatttc caggtgcagt gtgaagaaaa ggctgagggt ctgaggcagaaggggagggc 4260 aagaggctgg gcccagtgct catggtccag ctggggctac catggaggccaggccagggc 4320 cgttagcctt ggatccattt gggggcttcc tttttgattt cctcagtccttgagtaagcc 4380 aagtggtact tctctagcca aaggcagagc cttgggcagg ctgcgccttgagaaacagaa 4440 ttctgaggaa gtaggctaca gctgggagag atggcaagga gctgggggttcactcccttg 4500 gacgtttcta agagggacat gtcactcccc tgggtgctgc tgtgtaggggtaaatccttg 4560 gaggctgggg aggaggcaca gggaggaaag ccctcagcag caggtgggcaagaagtctct 4620 aggacacagc caagtcagga gagggagccg ggctggcctc cctccatcggcatctcctcc 4680 ccccagccct gctctgccca ccttcctgga gtctctgtgg ccaggcctgggccagagagg 4740 gccaaggctg gcgtctcctg gttggcctag cacctggaca agtacaggcctcccgagcct 4800 gggatcaggg gatggggtgc attctagcag acttcggagc tggggaaggtgtggactcct 4860 tggggatctc agctctggtc cccccagggg caaagagggc tgaaaatagaacatagatca 4920 aagggtaaga tagatgagtt gatgagaata tgactggggg agagattagggaagggaacg 4980 aaagagctga caaggagagt aacagtcata atataagtta aggttggaaaggaccacaga 5040 gaactgcagc tccaaacaca tcctttcaca ggcaaggaca ctgtgggccaaagatcaggg 5100 agctctgcct aagacgtact agtctagtta gagacatagc tagttatggccatcccagaa 5160 ctaagaaccc agtcctcctg gttttggttc tgggggcctt gtactcctcaagaagttctg 5220 gagagagagg aaagagcgag agagggaggg agagtactgg gaaagtagctgtcacatgat 5280 tggcccaggc ctggcatttc tcaagatgga tgtctcctgg cctgccttggtccctcaaag 5340 tggtaagtgg tgatgaggtg tgagagcccc cagcagggct gtttgctcagcctctctgca 5400 gtcttggtgg tgtggtcagc agcgctgagc ctggctgggc gtacctagacagaggccagg 5460 ctgacagtgt gcaggctggg ctgctctggt ggaacagcag gcttgagaggcttggggtaa 5520 gaaaagggcc ctgggtgttg tggggctgga tcaggggccc tttgacattcatatgagaat 5580 aggaagagga ctgggctgca gcaaataacc tttgagaaca tgtcctgattgtgtagatag 5640 aaggtggcag aacaggtgga ggcctcagtc tgtcccactg caaagaaccatgtgtgtaac 5700 tattacacct atcccacatg ctttggagga gggaggggct ttgctttggtgatgctgggg 5760 gactgactgg gtgaatctgg cttcctatcc cttctgttgc ccacccccaccagcccctgc 5820 ccagtgatag ctcctgctcc agggcagccg ggcaagcagc tcggcattgctcagacttac 5880 tcatggaaaa ctttcttggt tggaaacaac agcaggagga cttcagggtactcgaggaag 5940 ccaagcaagg acctggctgc agacggaagg acatgctttc ctggctgggattgctggctc 6000 aataagacaa agtgatgcta ggttcctggc actcctgaag ccggatatgttagctcaagg 6060 atggaaggat ttgggcatgg ctctggaaag ttggggtcat gagaaggcaagtgggctaca 6120 gcctggaaaa tttggaggat gggaaactgg gggtggtggt gctgagatttgggggatgta 6180 gaaatgaggg aagaatcgga aacaaaggag gtgaagatag cagaaatgcaggatacttgt 6240 agaatcctta acatgcgtaa gtaccctgtc tcatttaatt atttaatcttccaaacccta 6300 ggttactgtt atgaccatta ttcatataga aaaactgggg ttcaaagaggaaatttacct 6360 aagttcccat ggctagtgag gtgccagagc caggtctgaa agccaggtatctgagtctgg 6420 gtccatactg cttgtccaca gaaagagaag tgtgggaaga ctgtcaaggatttgtcatcg 6480 tcaccatctt tctccgtcaa tatctccaaa tccatctctg gcctagtctccaagataggc 6540 aggcattctt ctttttcaaa atatcaggct cccacttgca caggctgaggagccacatgc 6600 aaatccagag accacagcaa gtgctaccct caaagctgtg ggtgtgcgtgtgtggctgaa 6660 gagcagacct gcactaaagg gcagagggga agcaggagaa ggcacagccgagagaagagg 6720 tgagctgatg atgctcacat ggtgtgttag ttggagcttc atagctagggtctggaagat 6780 tctggtgtta atcagaaggg ccaaagatca aaacatggta atgaaccatcctggggactc 6840 aaaggcttgg agaggagagc ttagagatag ggagagaggg ccaacttaggcaaggaaagg 6900 gtagaggaat gtaccagaac ctgtgttgag gaatattctg cagttattctttttcacctg 6960 gaatttagaa tgtctggcta gagaagccag gtggaaagta gtatggagctgggaatgggt 7020 atggggagtg tcaacatgca tgcatgccaa gtgctgacca gtgagcggaggagaggccag 7080 agtgggagca gagaggagct ctgggaccct ctccagggga atcctgagtggaatgagaga 7140 tggtcacttt tctggctaaa gacctctggg gacagaatat gggttaggacagagaagggg 7200 gaaggctgga tgagtggaag ccgttgcagg aagatttact gtccccgttcccatcactgc 7260 ttaccctctc cacctgcagc tctgccaccc cctcccatat ttattgagtgcctactatgt 7320 gcttttgata cacgagtcag ggggttgagg gagaccaaaa tttatgtcctccaggggata 7380 actttctagt gaggggagac agacaataca caataaacat agtaaataggtaaattacac 7440 agtatgtcag aaatacagag cagggtcgta aatgagagaa gggagaaactgaactgaggc 7500 aggaaagagg agacaaaagg gaggtggggt gggagcttgt ggcataggatggagatttgg 7560 attttctctg gttggaggga aatgctggca gtgcagaaat tggaagtctgggtttggggg 7620 agtggcaggg agacacagct gcccagctat gggagaaaaa tggggcaatctggggccagc 7680 tggggaggcc cacccagaga gcatgttcca ggccagccct tcaggagtgagcagtgccga 7740 cccagagcag gaacacagaa tcctgccggc ccctcctggg cccagctgtcccgtcactca 7800 cgcccgctgc ccattggtta tttttgctac ggaatgtgcc agccccttggattctcctgg 7860 ggaacagggg ctcaagttac cccctctcat cactcagctc cccatctgtgagtgggtgtg 7920 tttaggggtg tacatggaaa gtgcccatgg gtgtccaggg tcctctggctggaacgagtg 7980 tggacacaca tatgtgccct ctcagcacac gtccctgtgc atgtgtctgtacttgtaatt 8040 tgctttgatg ctctaggaaa caagaacacc tgtatgcacc cagaatgtacaagacatgac 8100 ctaaacttta gaataaaaga gcagccaggt gcggtggctc atgcctatcatcccagcatt 8160 ttggaatgcc gaggcaggag aatagcttga gcccaggagt tcgagaccagcttgtgtaag 8220 ataataagac ttcatctcta cttaatattt ttttaaaaat tagctgggcatgatggcatg 8280 cacctgtagt ctcaggtaca aggggggctg aggtgggggg aatctcttgagcccaggtgg 8340 tcaaggttgc agtgagccat gatggcaccg ctgcactcta gcctggacaacagagtgaga 8400 cctagtctct aaaacaataa agaaatctaa aataagataa aggggccacagactcaaata 8460 actgcagggg tcaagtagaa actgaaaatg agggaggtgc gctgagctcaggcatgattg 8520 ctccagccca aacattgtgc aggctgaaca aaactcatca gtctgtccctgtatttattt 8580 tattttattt atttatttat ttatttattt atttatttaa agcagagtcttgctctgtca 8640 cccaggttgg agtacagtgg tgcgatctct gctcactaca acctcgcctcctgggcataa 8700 gtgattctca tgcatctgcc taggttggaa ttctggctcc ctcaatttattggccatgtg 8760 accttgggca agtaacctct ctgtgtctca gtcttcctct tgtcgtgaggattaaatgag 8820 ctcattcaca tagagtgctt agcacaatgc ctggtacata ccaaacacgcaataactgtt 8880 aatagttact tagtagtcac agctcagaaa tcaggattgg cactacctgtgctcactggg 8940 atgataattc ccatctccag gacacatgga gtcctagaac ctgtatggctttgactaagt 9000 gataggactt ctctgagact cagtcacttg tcagttaaat gggtataggattattcataa 9060 gggtttccag ctcatggggt tattgtaaat gaggtaacat ttgtaaagtgcctggcactt 9120 agtaactgct aaacaaacat agctataatg gtcatgggga ggatagaattttgtgtatgc 9180 aaagtgtgca tagtgctgaa tagggcaggg atcgggtgga acacacaaaatatttggtga 9240 gtgtgcttcc ctgtgattga gaacactggc caactttgtg aataatgggggtacctctgt 9300 gcacctttgc ttgtgtgtgt gaatgtacgg gggtaggggg ccgcgcatgggacaatcgcg 9360 aggtaaggca agataaagcc ctctctggct ttcttgagaa gccttagggttttcacagtc 9420 tgagtccatg ttaacacgca gtccacaccc gccaggaccc ttgccctgcgtttgacctag 9480 gcgcccccac ccggcgctgt gcccttcggc gagttcggtt ctgcctggcacagtgtgtgt 9540 gcgcgcatgt ttgtggaatg agcaagtcga gatgctgctg accttccagagaggccccgc 9600 gggaggaggt gagggtggga ggaggctgcg ctgggctgcc agaaagtggcctgagctaga 9660 ggccattgca cccctttctg tgcctcagtt tcctcatgtg cctagggctccggaggcaga 9720 tgcagggtgg gggtccgtgg ctctcggcgt taggaggtga ccggtggtcgtgtagggagg 9780 caggtgaggg cctccccggg ggaagtaggg ggacaggaca aggaaggggccccaggtggg 9840 gtgcaggctg gcgagggagg ggcggactcc agcgccgccg ccgccgctgctgccgccgcc 9900 gtcgccgcct tacccccacc cggctcccga ggccccaggc tccttccgccacccgcgccg 9960 gctcccgccc gctccccagc tcgcccccgg ccccgcctcc gactccgccccgcccccgcc 10020 cgtcccctcc tcgcccggcc gccggcccgg ccccctcccc cgccatgaagaagctgtggg 10080 tgaagaagcg tttccaggtg agggctccgg gggcgggcgg cgccgggagggggcagggag 10140 gcctgggcgc cccggaggga gggcgggtca ccgcagctgg gcccggtggagggggcgctg 10200 gatcggcgcc tgccccaccc gagccccgcc gagggcggcg ggccgggcgcgatctagggg 10260 cgcccgggtc tgtgtcctga gcgcgcaggc ccctccccgc gctgaagggcagatcccccg 10320 ctccccgatc gcccgcaatc cccgcgacca ccggggaagg cccccgctgcagcgttcggc 10380 gtggagcgcc cacttgcttc tttgccacat cttctctctc ctctgtccaacctcaacccc 10440 gggccccggc cccccgcccg gcctgcccca gccccccact ctgaagctttccatcttccg 10500 gagctcctga aagcaacgca cacggatccg cgctgagctc aatacattccccaagccctg 10560 ccctgtcctc cgcgcacctg ggcctcctgc attcgtgagg ctctggccctctgcccccat 10620 catccctccc ccaccccctt gacacctacc caggtttctg tcctccccccagcaggaatc 10680 tgtgccctcg ttcctccatc tcccagtcac tcaagcaccc cgccccccgcatccccatct 10740 tctccttctc ctcactgagc ctgactacct cgtcttctat cctctttctctcgggctgtt 10800 gccctggtcc cgggctgttg ccctggtccc gggctgtgag cctctgtccagtgggatgtt 10860 ccagcttcca cctgcagcct gcagttctct ccttcctccc ctgctcctcccaagagccca 10920 gctgggtacc ctggaagaag gcaggggagt cattccctac aggggaagctggtcactctg 10980 ggtctctgcc cacctcccct ctcacacaca cactggccag ggaatgagtttctgcttgat 11040 gttgcaggtc tggatgggag gcacagggac cttaggaaca agcctccccctcaactactc 11100 attctggctt ttctctttca gaaaaccggc cattcccgcc gggcctttggccgactcacc 11160 catggtgcgt ggaccgtggg cgtccttgct ctagcccatg cctactcctcctcttggtcc 11220 ctgtccctct gtgaggcatc gagttcctga agacagccca tgagatgtggaaccctccca 11280 ctcaccccca cacttatcta ccacccaccc gaccaggccc cctgtgccctacagctgaga 11340 gaggacccag cagaagggag ggcggctcac tagcacaccc ctgcatggactgggtgccct 11400 gttctccatg tgaggcctaa tgggaaggag ttcattgcca tgctttggcaaccagtacgt 11460 ggctcctgct tgtcatggca gccagaggga aactgaggca cagaacctgctagaatctgg 11520 gaaagttgaa aatactccca ggaacctttt ctcctaacct aaccactgggcatttttgag 11580 gacgattcaa cagtagaagg gagggacctt gaggaaggtg cctgtcacatcatgatgcag 11640 acagataagg ggttggtttg caaagagggg tcaaagcaca atgcaaatattgtaatagag 11700 ggtgggcctg actcctaatg ggaggcccag gtctgcggct ggactggacacaagcaggtg 11760 tgtgtgtgtg tgtgtgcatg tgtgtgtgtg gccagtggca gcaccagtaagtgccaagga 11820 taccagaacc actggggcag ctggaataac aagcccaagt atgggggtcccccgtgctgg 11880 gcacatccca ggtatctccc tccccaccca ttgccacagg acacctctggggactgggtg 11940 cctcacgccc cttctgtctt gactgccctc catgccctgc cccacaaacgctctgataac 12000 agtctgtccc tgtctctctc ctgctgctcc tatggaagcg aagttttccgctcctgcaga 12060 aagcaaagtt acggtaggaa actggctcct gctctagccc cccgcatccccccctttccc 12120 acccggcccc ggcctctcct caccctgcct cagctgcacc cgatgccttgcagctggttt 12180 ggggtagagg acaggctggc cccgcggttg gtcgagtgcc ctggcagtacgactctgagg 12240 tgactcctct ttgttcctgg ggtactggaa cccagacttt agagccttggaacctaggac 12300 ctgatgattt tggggctgca caggggctta agctttcact gacaaggggaggagggagaa 12360 gggaggaggc tctgataatc cattaagata ttggcaggcg gggagggggtggcagtttgg 12420 agggccctac ggaggtaaac gtgagtaacc aggggcccag agatggagccagggcactgg 12480 catgggaggg gttatcctga gcagcccagg ctgggcaggg gatgtggggagcaaaagaga 12540 ggaggtgctg gcagccctgc cagtgataag atggagcctg ctgttggcagggaggcagaa 12600 ggcaataggg aagagttgga ggcagaggga ggagggccct gcccacacagaccccttctt 12660 ctccagactc agagacggct gaggatgaca tcagcgatgt gcagggaacccagcgcctgg 12720 agcttcggga tgacggggcc ttcagcaccc ccacgggtga gctcctggggtgtacaaaga 12780 gcaggcaggc gggttttcca taaggggtgc ctcagtctca cggtgctcctttctctaggg 12840 ggttctgaca ccctggtggg cacctccctg gacacacccc cgacctccgtgacaggcacc 12900 tcagaggagc aagtgagctg gtggggcagc gggcagacgg tcctggagcaggaagcgggc 12960 agtgggggtg gcacccgccg cctcccgggc agcccaaggc aagcacaggcaaccggggcc 13020 gggccacggc acctgggggt ggagccgctg gtgcgggcat ctcgagctaatctggtgggc 13080 gcaagctggg ggtcagagga tagcctttcc gtggccagtg acctgtacggcagcgcattc 13140 agcctgtaca gaggacgggc gctctctatc cacgtgtaag taacggccttacctgggcct 13200 gaactgcccc atctcaccac gctgtcctgc gctgccctca ctgctcagtcagcctccacc 13260 catcaccctg ccccatccat ctctctgtgc atttcttcac cccctgctgccactccatct 13320 tcccacactg ctccctcctc ctcctgagcc atcaccgccc acatccccctgctcccacct 13380 gtcctggctc accatgccat ctccatggtc tcctggaccc tgctgtcccttccttgtctc 13440 ctccaagatc tccagtttct caggggccct cttttgcctc acccatttgggctccagttg 13500 tccccaggat ccctcccccg acccgggggc ccccttggtg cctgctgtctcagcagctgc 13560 tgccttttca tctctctgca cattcctgtt cccatgtggg cctttttctgggaggaacag 13620 aacctttcca cacggcagct cccgggagag caggagagag caggggaacaagccagcaag 13680 caggagagag aagagagtga ggtggccagg ggcagatggg gcaaggggcctgtgaaagca 13740 ggaggccatg ggctgggggt ggcagggggc tgggaaaggg aggggctggaaatggggcca 13800 ggccagaggg agagggcggg agtgatggtg gcagggggct tgcaatgatttctctcatgg 13860 gaaaccccta agtccctgag ggtgggattc aaggttgtcc caggagggggtgtgaggagc 13920 ggaggtgttg gaggcactgg agccattttt ggagatttgg ggctcgcagatacaggaggg 13980 agtgctatag tggaagaggg gagtggctga gaatagaggg actggggcatttggggagct 14040 gaggggggcc tggtttgtga tgggtatggg tgtgggggca gctaccaccgtgcaggagaa 14100 agaaggggcc tggtggggga ggctgctttg agggtggtta gagcagggctagcggtgggc 14160 aggggagagg gccagggctg ggccacggcc agggggaggc tcccttggcttatcttcttg 14220 gccttacccg gtgttcctgt gccctggact tgcctttccc ttcctgcctttctttaccag 14280 gccccagcca gagcctggag ttgctatggc aacttcggag gaacccatttacttagtgat 14340 gtctatggta cagagactgg cctggagctc agagctgccg gcaagaggcctcctctgctg 14400 tcctcaattt ctctccaggc ctcacccact tcccagaggc tcttccctggggactctgcg 14460 gcccttcccc cagagaagac acttcctccc tgcaggtggc tggctggggtttctgtctct 14520 caggccactc tgcattgcca ccaccccttt tagccccagt cagaggtgggtctgtcatgt 14580 gggtggctga ctcaggtagg ctaaaacatc cttaactcgg gacccctagaactgctcatg 14640 gctggcaccc tactacttgt cctcccttct gtgcccctgg aaacccagacactttggagg 14700 aaataagggc tcaggattct gactgcctgg gtttcaatcc tagcaccaatcatttcccag 14760 ctgtgtcacc ttggtaaggc atttaatctt ttttatgcct cagtttcttcatctgtaaat 14820 gggggtgatc acagttctta cctcacaggg ctgttgtgag tattaaatgactcaatgcat 14880 tttaagcact tggtacaagc ctggcactca ggaaatattc aatgagccattttacagatt 14940 tttattaagc tctctctgat gtgtcaggta tgataataca tttaattccttttttttttt 15000 tttttgctta tctttatgtg tgatgtgccc cctcacccac cccctcctccccgcaagggt 15060 ccagatgggg aaaactgaga cccagctctt gggcaccaaa gctctgttaagtgaaaggga 15120 tactctgggg tgaccggctc cttccctcct ccttttctcc cacactccctattcaggccc 15180 acttagtagc tatttctgag ctgagttatt tcagagcata tccctgtgggggggggcctt 15240 ctgtacttct caggggggat ttctaaggac tcaaggtagc tttgccaggggaagcacaag 15300 tcaaaggcct atcggggggc agactgagca aggagagtag gagcctgggcatcccgttgc 15360 cacctgctgt gtcccatcag tgctcggggt ggcggcaggt ataggctcaggtctacacag 15420 cagctaggga atcctaggga tggggcactg ccccccaaaa ggcttgtgcctggtccagga 15480 ggctgttgct tggcttccag ggaccactag gaaggggtgt gccctgctgatgatgggcag 15540 gggtgtgggg gccagctggg ggctggaatg agtgggtggc tgcattcctgagaacgcccc 15600 tccccacccc accctcttgt cttccctccc cacttcatcc tttgggtcctaagcctcatt 15660 cttttctctc cctcatgctc agttctgttt ccgctgtttc tcggcttccagggctggggg 15720 gaggaggctg gcccgagtcc tggggctgag tctgtaccaa gacccagccattagcccaat 15780 cttgtggttc cagagccgcc ggcctctccc cagcacctgc tctggctgtgcgctcctcgt 15840 gggggtgggg gtggggggca ggaggatctg gcccatgtca cccccaagcctgcccagcat 15900 gcccaccacc caattcctgt cacaagctaa gggtctagga gaggaggccccctgaatcct 15960 ctacccttct ccatcttggt tctgcagcag cgtccctcag agcgggttgcgcagggagga 16020 gcccgacctt cagcctcaac tggccagcga agccccacgc cgccctgcccagccgcctcc 16080 ttccaaatcc gcgctgctcc ccccaccgtc ccctcgggtc gggaagcggtccccgccggg 16140 acccccggcc cagcccgcgg ccacccccac gtcgccccac cgtcgcactcaggagcctgt 16200 gctgcccgag gacaccacca ccgaagagaa gcgagggaag aagtccaagtcgtccgggcc 16260 ctccctggcg ggcaccgcgg aatcccgacc ccagacgcca ctgagcgaggcctcaggccg 16320 cctgtcggcg ttgggccgat cgcctaggct ggtgcgcgcc ggctcccgcatcctggacaa 16380 gctgcagttc ttcgaggagc gacggcgcag cctggagcgc agcgactcgccgccggcgcc 16440 cctgcggccc tgggtgcccc tgcgcaaggc ccgctctctg gagcagcccaagtcggagcg 16500 cggcgcaccg tggggcaccc ccggggcctc gcaggaagaa ctgcgggcgccaggcagcgt 16560 ggccgagcgg cgccgcctgt tccagcagaa agcggcctcg ctggacgagcgcacgcgtca 16620 gcgcagcccg gcctcagacc tcgagctgcg cttcgcccag gagctgggccgcatccgccg 16680 ctccacgtcg cgggaggagc tggtgcgctc gcacgagtcc ctgcgcgccacgctgcagcg 16740 tgccccatcc cctcgagagc ccggcgagcc cccgctcttc tctcggccctccacccccaa 16800 gacatcgcgg gccgtgagcc ccgccgccgc ccagccgccc tctccgagcagcgcggagaa 16860 gccgggggac gagcctggga ggcccaggag ccgcgggccg gcgggcaggacagagccggg 16920 ggaaggcccg cagcaggagg ttaggcgtcg ggaccaattc ccgctgacccggagcagagc 16980 catccaggag tgcaggagcc ctgtgccgcc ccccgccgcc gatcccccagaggccaggac 17040 gaaagcaccc cccggtcgga agcgggagcc cccggcgcag gccgtgcgcttcctgccctg 17100 ggccacgccg ggcctggagg gcgctgctgt accccagacc ttggagaagaacagggcggg 17160 gcctgaggca gagaagaggc ttcgcagagg gccggaggag gacggtccctgggggccctg 17220 ggaccgccga ggggcccgca gccagggcaa aggtcgccgg gcccggcccacctcccctga 17280 gctcggtaag gcctcaggga gggctgacaa ggtgcctgaa cccccgtcggggggcgtttg 17340 tggagagcaa gactgctcag caggagccgg ggggtcgggg gtttcgcctggggctgctag 17400 ccagctgcaa gggtgggttt gccaaagaag gcacagacac aggctcgactttgagtgaag 17460 gattgtcaaa gtccttgtgc taggactgct actggtgagg cagagcgtgagtgttgtgat 17520 ggaggctagg tgaggctgag atgtgagcta agactggtgc ccagatgcccgccatagctc 17580 ccctgggtcc agtggcctgc taggctgttg gatcaagaac cactactggggggatgcact 17640 ggtgggaacc taaggacccc cctccatacc cccaatccct ctgttggggagagatggtag 17700 atggtctgaa ataattttca aatccctgtt acagacacgc tttgtgccagataactcttt 17760 actctgcctc tcccacccgg gcaccatccc ctgacccatg tgtggccacccagcctgccc 17820 ttctagcctc ctcacctcag gcttacaccc cgcatggctg agctcactgtggtccctaca 17880 caatgcctgc ctgtgtgttg tctcccaacc ttcccatgga tgatgaccaacaccaccaac 17940 aggagaatgg ctctgcaccc cacccctcat cctgcacatc aactcgaggcccactcctgg 18000 agagaggcag aggaaggcgt ccttgaccca cttccactgc tcctccagaatagatgcctc 18060 tacctgctgc tcctgggaga ccctgccagg atctctttca ttgcacttaccatactgtat 18120 ttttcttcat caaaattatt ttgcatttaa gtattaccac cttagaatctttagagataa 18180 ttaggcccct gtccatcatc ctcctggaca ccattattgc aaacacacaactatgtgcca 18240 gcattgtgtt aactaggccc ttacagatgt tatttcattt cagcctatatgtcagtttca 18300 gagggtcttt tcccctccat cttacagata acaaaactga ggctcagagaggctaatttg 18360 cccaaggttt tgtagctagg aaacagcaga attgggattt tcactgcttttttggctatg 18420 tacattgtcc tttatctagc ttatggaatg tcagagttgg agaatgcccagagacccatg 18480 acagtggagt ttgtcatctc tctgtatgta tgagcttcct gtaagcagggaccactggtc 18540 cttcctctag tgttcttggt acccatacag tactcagtat gccgggagtacagggtcact 18600 atttggtgaa cggatgaaat caaatctgat cttcgtaggc ctctcagactgcctaccatt 18660 cactcttttg aatctgacgg cttcacacat ttgacaattc acctcgtgttgctctgtgac 18720 accgctgtta ttgcttaagt cttgggttgc ttttagcttg ttctttgtctgtttgaaagc 18780 ttgtgcccct ccaggtgcct tgcatagggc ctggtacgcc atacatgatggctgacgtgt 18840 taaagacata cacagtgcag tcctgtcttt ctccagaagc caacgctctaattgtggatg 18900 aagttgagga acggccacac taacatggag tcaaggctgc cctgacctgtttcgagaatg 18960 ctccttctga cttccttttt tccctaagga cattcaggag gcagaccctcttctccccaa 19020 gtccctgctt tctagaagcc cctctgtctg ggtttggctt tctaggatgcaggccaccag 19080 gactctctct cccgctgtca tccctgcagg gatcatggcc ccttaccccgttattcctgt 19140 gtccggcaga gtcttcggat gactcctacg tgtccgctgg agaagagcccctagaggccc 19200 ctgtgtttga gatccccctg cagaatgtgg tggtggcacc aggggcagatgtgctgctca 19260 agtgtatcat cactgccaac cccccgcccc aaggtgagct ccagcactgggccaaggtgc 19320 ggtcgaggtt gggagggggt gtgtgagaag ggaaggggag gttcccccggactcctccaa 19380 gggaggggtg ggaaagaggg gaattatccc ctccacgggg gctgccctgacttgggtgtg 19440 tgttcaggga catttctcag gacccccgag agaagggagg gcaacctgagcttcccaaaa 19500 tgagggcgga ctcttccaga ttccctgggg tgctgagagg agaggtttggtctcctgtgt 19560 ggtgtgtggg gtaggagtag agattctcag tgggcgcctg tgggccgtggcgagccgggt 19620 ccctgtgcct ccccacagtg tcctggcaca aggatgggtc agcgctgcgcagcgagggcc 19680 gcctcctcct ccgggctgag ggtgagcggc acaccctgct gctcagggaggccagggcag 19740 cagatgccgg gagctatatg gccaccgcca ccaacgagct gggccaggccacctgtgccg 19800 cctcactgac cgtgagaccc ggtagggagc ccatcaaccc tggggctgggtgggggcaag 19860 ccgtgactct tccctggccc aggccccagt ccacctccct tcccactctcagccttgagc 19920 ttgggcaccc cgccagcata cttagtccat gcagtccctt ctgggtgtcggcagctttgg 19980 tgaaagcgtt ttaaatggcc ctggcctcag ggcaggggcc aaatccccaaggcgcacaga 20040 aggctggcca attcatgaag tcagtgaaat ttgtgtttag ccatcctctaaatgccatcc 20100 tcccatggca tttccctgaa cagggtccta tggggaaagc aaattgtccttgagtttccg 20160 agaagaagaa tctctccgct cacagggttt aagagccacc acaagccttctgaagtcatt 20220 tcccctgtaa gacagtcccc cctcccagta aaaagagatg ctttcgagtgccacaagcca 20280 cttcccccct acttttcttg aacccaaagt tgtaaaaagg gtacagccaagagcacttct 20340 tgggctggca gtgccgaggc tgccttgttt tctatttttg tgaaagcccttacttggtgt 20400 cagactactt tctttgggat tcagcccagt ttctttctgg ttcagctggatcagtgtcgg 20460 tgtacatggc cagttcagct cattcagctt gcggggcctg acaatgcactgaccccgggc 20520 ctggggttcg gggaggtgag gatggtcagg gggtattaca gaaggaaaacacactcaacc 20580 ctcaaggggg ttcccactgg gtgaccagac ctggatcaga ggacctaggttgggggacag 20640 tgcggagcag atggatctag tgccgaggac atccgagccg ttgcttagtgttctgggatg 20700 cctgcgctga gagacgttgg tgccctgaag gaggctggga ttttagaccggcacggggaa 20760 ggcgggacat gccaagaggg ggaacagcac atggtgttct gtcccttccttccatgaagt 20820 gctgcaggag acaagatggc agagcctgtg tgcccatccc agggcctcagcacctgagca 20880 gtgagggagt tcttgatgca tctgatcata gtctcgtgct ccagtagagcctttggttgc 20940 acggtctggc ttgtgccact gcaggtcttc atcctcctta gctgcatatccctcccgggg 21000 cccattttcc aggctttctc ctcctcctac ccctcctacc ccgctgaatcatgcccgtcc 21060 ctccaccaca tgcttctgtc cttccatcac ctccgggatc tggcttctgactcagctccc 21120 agctcccctg agggggccca ggcctggctc caggacccca gtcaatacctggcttgggct 21180 gaaatttggc gaggggtggg atgggggtgc cccgatactg gctcagggccatttgggagc 21240 cttttatggt tggaaaggca gcttggggcg aggcagttgt cagcccttgactgagagttc 21300 tgtatttgcg gcatagaacc tcctgagctt cagtttcctc atttgtaaaccggagataat 21360 gacaccgtcc tcacggatga aagcgaatgt gtgaacgagc tttatgaactgtaaagctgt 21420 agacaaatgt tagttgttag cgttattaac gggtgctgtt gtgtagaagagctcaggttt 21480 ctacaaggat ctgggaagtg ccttatcctt ctctgtccct ttcttcagcccatctgtgcc 21540 ttagggactc cacctctccc cttggaaggc ccatatcctg cagaccctctgtatttccca 21600 gcccctgtgt cctcagctca tcacagcatc tcggcacctc tgccctggggagccagaaag 21660 ccttcattgc attagtctgt tgcatcaggc attacagcaa gacagccacctccttaagtc 21720 aggctggctc gggggcccca gggcctgggg ggcaggcatg tgggtccagacttggctctg 21780 gctgttgtgg agcaagctga ctttcttacc ctacaaggca ctgtttagtccagagtggct 21840 ggatgggggc cagtggactt gagagcagca aaagtgggtg gaacctggggatggcacaga 21900 catctggcaa ggggtggtcc caggcctgga gtctgcaggc agtgaggggtggggccaggg 21960 gagggagtcc agcagtttgc ccgtaggcct ggtggggagc aggtagagggaggcagtggg 22020 cagtgtattg tgggaagagg cctgcgtgca gccaccgccc tctcttgctccttcccctcc 22080 ccctcttgct cttcacgctg cccctgccca gtgctatggt aaccagggctggctgcccag 22140 ttccctcttg gggtccaccc caacagggcc tgcttttgag gacccacagatctcaattcc 22200 tggttgggag ccatggtaac cagggaatgg ataactagag aatgcggccccatggaccat 22260 ggtttagggg cagggtctgt gcggaaagga gctggtcctc accagctcccctggtccctc 22320 ccaccctttc caccccactg aatcattctg ttgagaccac agccattctcccaaagcgga 22380 tgtgtggatg ggggcaggtg gcttgggagt gtgagaagtg tgcttaaccaaagcattccc 22440 caccgctgcc cacatccatc acacaagtat ttattgagtg ccaaccaggtgccaagtgct 22500 acagctctct aaacagtttc ttctgtttgg acagtgtcat agactttcccaagccctttc 22560 acgtctcttc cctggcgata acattgtgcc atacatatcc cttggagggacgtgtggcag 22620 gtggggacat tcccatttta tggatgagaa gacaaaaatg tgaattgaaaagtgaggtag 22680 agctagcacc aggccagcag cctttattta gcacttgcca tctgcctagggatgttttca 22740 gcaacaactg atttagggtg atggaaataa ctgcccatga agcaaataaacttagtgtgg 22800 ggtgcaaata aaagtaacat gactgggctg agtgtggtga atcacacttgtaatcccagc 22860 actttgggag gctgaggtgg gtggatcacc tgaggtcagg agttcgagaccagcctggcc 22920 aacatggcaa aaccttgtct ctactaaaaa tacaaaaatt agccaggcgtggtggcatat 22980 gcctataatc ccagctactc tgggggctga aacaggagaa tcacttgaacccaggaggcc 23040 gaggttgcag taagccgaga tcgtgccact gcactccagc ctgggtgataaagtgagact 23100 ctgtctcaaa ataaataaat aaataaataa ataaaaaata catgaccaggtagcacttat 23160 tgagtgcttc atatatacca ggccctgtgt tgattggatt attttgtttaatccagcctt 23220 atgaggtagg ttctattatt atgcctatgt tacgaatgag gaaactgagacttggaaaac 23280 tttagccaat tgctctctgg acacagtggc tgacacctgt aatcccagcagttttgtctg 23340 gagcagcatt gatcaataga catttctgtg atgatggaga tgctctatatgtgtgtggtc 23400 cagtatggta gccctagcta catgtggtta gtgagcactt gaaatgcagctgcggtgact 23460 gaggaactgc attttaaatt taatttcatt ttaataactg cttttaaatagtctcaggtg 23520 gctagtggct gccttcttgg gatggtcagc tctaggagct tagggccagctggaggctgg 23580 agctatgatt tgaaaccctg tctcggtctc tcaagcctac actcctaaccatgacactat 23640 cccacccccc tttcctgttg ctctgaagaa gttcgaggtt gaggtagaactgagctcatg 23700 agctcactgg gagggcactg gtgatgccct ctgtgggtgg gtccgactgggggctttctc 23760 ttcttcctct attgactccc cacctccagc acagctgtgg tccttctgtttccttgcatg 23820 cctgatgtga gcacacccag cttccccacc gtgcgccccg agaggaggccctctgggtgc 23880 tgggcaggag gaagtgggag tatgggaggc cccatgggcc tgggcctcatcctccccacc 23940 tccagtcctt ccctcaacta caggtctcct atatttgagc ccaggattcttgcattttcc 24000 agccaagacc ttggcctctt cattgtacct tagctggtca tggttttcttggctgtaaag 24060 tgaggatctt cacaccagcc aggccaccta cttggtgaga ttcctgagggtctgtgagga 24120 gggcaagttg gacacttggg acagaggata tatagctcat cgaagcccattctggtccca 24180 ctcatagaat caacatgacc acagcagaaa ataggttgtt tattgggggccatgaaaaca 24240 aaaacaagga agaaaacctt agcatccatg tcagtccctg tcttctcttgcccctggtct 24300 ctggccacct gtggcatagt tgcagattct cccttgtgat ggcagtggctgccgttgggg 24360 aggatgccac atggccccca tcgtcagtcc tcgatggggg tttggacaacaccagacaaa 24420 gcagtgtggc atggtcccct tgcccaaact cagcccttcc tctaccctcctgtctgccag 24480 aaacaggcct gcctgacagg tggcctcccc gaagctggta cctagaacgggggccatcac 24540 tcacagcctg tcagaagaat tgagagctca tttactgttg ttgcagccacttttcctcac 24600 ctggagcaaa ctaacatagt catatataag aaattcatca tgaccgggtgcggtggctca 24660 cgcctgtaat cccagcactt tgggaggcca agcagggtgg atcatgaggtcaggagtatg 24720 agaccagcct gaccaacata gtgaaaccct gtctctacca aaaatacaaaaaattagccg 24780 ggtgtggtga tggacgtctc taattccagc tacttgggag gctgagacaggagaatcact 24840 tgaatctggg aggcagaggt ctcagtgagc tgagatcata acattgcactccggcctggg 24900 cgacagtgcg agactctgtc tcaaaaaaaa aaaaaaaaga aaagaaaaagaaaaaaaaga 24960 aaagaaaaaa gaaaaattaa tcacttctgg ggctaggtca ggctgaccatgtgaaatgat 25020 ctgttggctt gtacattttg aactaagcct gaaacattct cagtccaaggggaaacattc 25080 atgtcagcac cctgtggaaa tgcccacctc ctggccagcc gtgtcctccctttccctgac 25140 ttttccgcag tggggttacc acagcagaca tttcaagaac tgcctgtggaggacctactg 25200 aaacagcaca tgtggagggc cagctgggaa gggccaagcc acaggcaaatgcttgctaac 25260 ctgcagccta tcactccctt cctcaagaat cataaggagg aaaaaccgtttatggactgg 25320 gtgccgtggc tcacacctgt aatcctagca ctttgggagg ccaaggcaggtggaacactt 25380 gagaccagga gtttgagacc agcctggcca acataatgaa accccgtctctactaagaat 25440 acaaaaatta gccaggtgtg gttgtgggag cctgtaatcc cagctactgagggaggccga 25500 gccaggagaa tcgcttgaac ttcggaggtg gaggttgcag tgagctgagattacaccagt 25560 gctttccagc ctggatgaca gactgagact gtctcaaaac aaaagcaaaaacaaacaaac 25620 aaaagaattg taaagagtcc gccttgccta cagagtaaag gctgcacacctttgtgtggc 25680 ctctaccatc tgcacccata gggcaaaagg agccagaggc ctgaggctggccctcgggtg 25740 gcagtgggag gcacctgctt gaacagtgga tgctccactc cttacttctttacccagttt 25800 atagacagac agttgagcta tagtcctctg cacacatggc ttttgcttcaaacttttccg 25860 catgcacttt cctctgcctg gaatactttt ccctttttcc tgcaaactgcaattccaata 25920 ctgcctcttc cttcagttct ctctttttcc ctgggccaaa agtcatctcccgccttctat 25980 actcccacgc cccttacctg ctgctctctt gctacctcat acttcctgcttgctcttgta 26040 gtcacttgtg tatctggttt gtcttcccca ccacccagaa ggctcctgaggacatgacca 26100 tgtctttgcc actccttagc actggcagct gtgcctgggg gccctcctggacagttaggg 26160 gctgtgtgga ctgcagagct gtatgtgagg tggcagtgag taggcagagagccccatgac 26220 ttttcaagcc cttgagcaac gtgggtgaca cacagaggtg aggctaggcatgacacccag 26280 ccacaagggt catgaggctc tgcctatgag gggagcttgg caaagagaccatggagcggt 26340 ggtcctcaga gtgaggcccc cagaccagca gcatcagcat cacctgagtatgtgttagaa 26400 atgcagtttc tttttttttt tttcccaata agcgttttgc actcttaagaaatgcagatt 26460 attggcccca gccccgacct actgaatcgg aaattccgtg ggtggggcccagcactctat 26520 ggtctaagga gccctccagg tgattccgat gcacagaaaa ccttgagaaccactgccata 26580 gagttagaga ttgtgcccaa gatgggagac caggtcatgc caggggaagcacatgggctg 26640 tgtagccaaa cctcctaggt tcaaagctct gctctgaggt tgaatggcccagtgaagggg 26700 gcttagtgat accacctctt agggctgtcc tcctcccata aagatcagcagtggtcccta 26760 agctttcatg tgcctaagag tcatctgggg tgctttttaa aatgcagtttcctgacatct 26820 catttcagaa aatctaattc tgcaggctgg agtaaggttg aggaatctatattttaaata 26880 agcccgaggt ggtcctgatg tgagtaggtc aagtaacatt ctgagaagccctgaagcaaa 26940 gggctgtgtg cgacaccttt tatgtactag ataattcgca gtggtggctgcaactctgtc 27000 tattgtgaga acacatcagg ggacgctgaa acagttgggg tcggtggatgtgctggtctg 27060 aggagaggga gttccctgtg tgcttcctaa cgccagtatt ttcatcaaatgagtttgacc 27120 tgagagtgat gttgaaaggg cagacatgag catggggtca gcgtgggggattgggagcca 27180 gtgtgaaggg gcggtacttc cgtggtgggt tgttcggggc cattgagtgtatgtgcctat 27240 taggtttagg ggggcacctg ccccctcctt cctggctggt gtaactgtgtttgccacagt 27300 gagagttggg cttgtgggtg cagcagggct gggatgtcag ggactggttccaaagatggt 27360 ttttgctatg tggccaggga aaaaactagg tggcaccgga ttctggtatggagcctcagg 27420 aatcctcgat cagcttcctt ggtcactcac ttttggaggt atggaggggcagaggctact 27480 gccttgtctg gaagaggcct gggctgctct gacagtctgt ctctccaaggggcttgagga 27540 tgggggtctg cttcatggct agatgccctc cagcatgctg ttcccctgggcaccaccagg 27600 ggtctctgag gctccctgca aaccttgacc atctggcctt cagctctgcttccgttccca 27660 gtccctgggc ccgtgagcct cctcagtact gtccagcctg gaggtgaccctggggcagga 27720 ctccaggctc catagagggg taggaccgcc cacctgcgga gccatgcctgtgatctcagc 27780 atcaaatgcc ttaagcacca aatgccattc tgacttccct ccccaaccctacctcaagac 27840 agccagcctg aaccgtgggc ccctcctctg cccggccccc agccctccttccttactggc 27900 catgctggga aacacaggtc atggcttggg aatgtggccc cgggttgcggggtgaggtga 27960 taggaagagg gagaaggaca tgtgacccct gctcaacagc ccccttctctcaggttctgt 28020 gtccaaaccc tttccccatc ccagatacaa atgggttctc atgataaggggccatttggg 28080 ggaggcagcc cccgctgtcc tctttagcgg ggctaaggtg ggtggcgggcaagttgccaa 28140 caggtgccct ggcggtttgg gtaggaaggc tggatgtggg cttaggccctgtggtggctg 28200 ctggccagac cttccatggc agggatgagg gggcagaggt gggattctgggccccctcag 28260 attcttcccc cacctctgtg aaggagggca aagatcttga cagttctccgattttcgggg 28320 ccaaagaaaa caggtatgcc ctggctctgt agtatttgag gctcgttagcgcattccccg 28380 gtcagggatc ttggtggttc cgtcagagca aggggcaaca cagggaacatttcccacggg 28440 caccttcttt ggtggacgtg tcagaacaaa tgggtcaggg caagtgtccctattggcaca 28500 gcgcagtgcc acccctcccc tgtcttgctc ggggacggag gccccacaccttgagtcaag 28560 gcaccgcaga gcgggctttg ctcttgtcag ggtctgaagc tgtaaggagaagaaaggcag 28620 atcccgcggc ttgggagtga gcagagtggc taaatccaga cggccgcgctccgccctccc 28680 tcccctctcc cctctcctcc ccctccacac cagggcccag gctgcctgtggtctcggcag 28740 gcaccacctt cctagccagc tgcctgccgg cctgccatcc aacctcctccttcccctcct 28800 caggcgcctt cccctccccc agggccttct ccgtgcaggg cctcagctgggtcagccgag 28860 tgagtggggc tgggcaggct ggacaggctg ggctcctgct ccctggggagccacgctggg 28920 gtgggcagtg ggcaggcagc aggagggcct ggtgccaggg cagagccttcaggacaggca 28980 caggctgggg tctgtgtgca gagcctcggg gtgagttggg gtacagccctgctgggggct 29040 tggggtgggg ggaccctggg gagaacctag ggggctgtgg cagtttcatgtctgtatttg 29100 gcagtcggat aggagccagc tcagcgagac tgggaccctg gccagttgcaggggagggcg 29160 tcagggccct ggggacaccc ctcccccaag gctgactctg gtgcccccctcctcttcccc 29220 caggctgagc tgcccttttt ggtggatgac tccagctctg ctttcctatccctcgagcgt 29280 ttgggggtgc aagctgtagg gctgtggcca acaggtgccc ctggggtttgcacggcagga 29340 agctgagaag ggaaaggggg aggggcaggc tagatagtgc cgcctcattggccctggacc 29400 gaggtcggga tgtgacttgt ctgcggtgtg tgttcctctg caccaggcccagctcaccca 29460 agaaatgggc gtcctagcca tgctgctcca gggttccata gacctgcttcccctgctttt 29520 ggagcctgca gttggtagtt taggtctcta gtccatgtca ggaatgtgggtgccctgctc 29580 cagtggaatt ctcccaccag gcccaggagt ggcccctctg ctgcccgaaccctttgcccc 29640 aggcctttcc ctatggtgtc ccctcctggg aggatcagca gggtcggtgttggcagagcc 29700 agtggcagag gaggttccag gggctcaggg aggggaagaa gctgggcgaggtcgcaggag 29760 cctgggggtg aagtcagggc agggccggcc tgcggggagc tgccgcaactcctgggctct 29820 gggcgacatt ccagccagct ctcccaggct ctctgctcgc cttcctccctggtagctatc 29880 tctgtctctc tccaggtggg tctacatccc ctttcagcag ccccatcacctccgacgagg 29940 aatacctgag ccccccagag gagttcccag agcctgggga gacctggccgcgaaccccca 30000 ccatgaagcc cagtcccagc cagaaccgcc gttcttctga cactggctccaaggcacccc 30060 ccaccttcaa ggtcagaccc ctgaggctgg ggcctagcct cctgtgtgcccccgttcctt 30120 tgggtgcccc cttgttcttg gggccatgcc taggcaacat caagacctggaactttgctc 30180 ccattcaaac cctcttaccc agagccagtc tcagcctggc tgtggaagagtcctccgtct 30240 cagattccac agcccccagg acccaaggct ctgccctgtc ttcccctgacactttggggt 30300 acccccttca ggtctcactt atggaccagt cagtaagaga aggccaagatgtcatcatga 30360 gcatccgcgt gcagggggag cccaagcctg tggtctcctg gtgagtagccgcactttcca 30420 ccacccacca gcgactctat gccaggcctg gctctgggag gtctggctttgggtggaagg 30480 aaatggaatc ttggtgggct tccccatgat ctgcctcagg ggcctcatctcagggacagc 30540 agtgtactcc ccccggcacc ctgtcccttg ccccatgttc caggcttatttagggcttcc 30600 ccttctgggg aggggtttgg gtctcatgtc tgtccatttg ggataaatgactgggtgcgg 30660 tggctcacgc ctgtaatccc agcattttga gaggtgaggc gaatggatcacttgaggcca 30720 ggagtttgag accagcctgg ccagcatggc gaagccctgt ctctactaaaaatacaaaaa 30780 aattagccag atgtggtggt gcatgcttgt aatcccagct acttgggaggctgaggcaag 30840 aagattactt gaccctggga ggtggaagct gtagtgagct gagatcacaccattgcactc 30900 cagcctgggt gacagagtga gaccctgtct caaaaataaa aatacccccttcccaaaatt 30960 agcaaggagc aagacatttc agaggccaag gaaggaggat tgcttgagccaaggagttga 31020 agaccagctt gggcaacata gtgagactct gtctctacaa aaactttttttaaattagct 31080 ggacgtggta gtacatgcct gtagacccag ctacttatga gggtgaggaggaggatcact 31140 ggagcccagg agtttgaggt tgcagtgagc tatgatcata ccactgcactccagcctgga 31200 caatatagca agaccctatt gctgaaaaaa aaaagacatg caacaatttgtttctgagct 31260 gccagcagcc ccgagttaaa tgtgggtcta agcagagggg cctcgctcatcccaggtgcc 31320 tggaagatgg attactcagc ttgctaattt tttatggttt gaattctgtttttcattttt 31380 aattgattgt ctggtcccca ctgtgatttt tttttttttt tttttttttttttttttttt 31440 ttttagctgg agtttcactc tttgtcgccc aggctggagt gcagtggtgcgatctgggct 31500 cactgcaacc tccgcctcct gggttcaagg gattctcctg cctcagcctcctgagtagct 31560 gggattacag gcatgtgcca ccacacccag ctaattttgt attttttgtagaggcagggt 31620 ttctggtcag gctggtctcg aactcctgac ctcaggtgat ccacccacctcggcctccca 31680 aagtgctggg attacaggca tgaaccaccg tgcccagccc ccactgtgattttttttttt 31740 aacattgtaa gttttttcat atcccttttg ggaagtggga aagctatctgtccattataa 31800 ataaataaaa ataagacatg gggaagttta agaattatta agagctaaatagggtcaggc 31860 atgagggctc aatgtctgta atcccagcac tttgggaagc caagagaggaggatcatttg 31920 agcccagaag ttcaagacca gcgtggacaa catggtgaaa ccctgtatctacaaaaaata 31980 caaaaattag ctgggcatgg tggcgtgtgc ctgtagtccc agctactcaggaggctgaga 32040 tggggagatc gcttgagcct ggggaggtag aagctgcagt gagctgtgattgtgccactg 32100 cactccagcc tgggtgacag agggaaaccc tgtttaaaaa aaaaaaaaaaagacaagaaa 32160 aaagagctaa atagcacggc tccactctga atgcagcagg gttccgagaagggagagctg 32220 caggcagtta gaagtgacct ggaagctcct agaggtgggt gggatggcagagtgggggta 32280 aaaacctagc cctggaaacc agggatgccc acggtcagtg ggacagatctggggactggg 32340 caaactgcac agggaaggag aggcctgcac agtgctgcag ccccagttcctgtgcacgca 32400 catcaggccc ctgggccctg ggactgagtt cttgcccctc tgacaggctgagaaaccgcc 32460 agcccgtgcg cccagaccag cggcgctttg cggaggaggc tgagggtgggctgtgccggc 32520 tgcggatcct ggctgcagag cgtggcgatg ctggtttcta cacttgcaaagcggtcaatg 32580 agtatggtgc tcggcagtgc gaggcccgct tggaggtccg aggtgagtacctgatttctc 32640 catgaatgcc cacctggccc tggccccttc cttcccccac tgtctgctctcacacagcct 32700 cagttagagg atgccaccac tgaaagggcc ttaaggggcc cctagtccagctgcttatat 32760 tattgttgag tgaaccaaag cccagagaca ggaagtgacc tgcccaaagctgcacagcac 32820 attgttccgt gctcagctta ctacacaaca ccaccttccc tgttgctccatcacggagcc 32880 ctggcggcag ccagagaccc tgcacctgcc actggccaag ctctctctacactcttctga 32940 gcctttgtca ccctgctctg cccaggaccc tccctcatcc ccctccccctctcctctcgc 33000 ccctttgccc accctcccat cccattgctg tgcagaagct actgtgaggtagcggtgggg 33060 agagtctggt ggctggaccc gttcggaggg gcccttgggg tggcttaggcttggagcatc 33120 acagggaccc tagggtgcca gggtgagcac agctgtgacg tgtgaagaggcctgggcccc 33180 cagagcctgg ggcatatgtg caggggccct ctcaggcagc agaaatgccaggccctggct 33240 agggggccct cgagggccat gggttttcct ccccaaaggc cacaaccgtttatcttgcct 33300 tccacctcac ccttcgcctc aatggctcct cgcttcctct cctcgcttctcactcagccc 33360 ccagcccctg accttcccca aggctcagag ctcagaccct gacagtcatggtccctctgc 33420 tggcccccct gcctcagttc ccctcctctg tgcctgccgt tcctgtagttgccacttcct 33480 tggtctccag attcttcagc cctcctcccc ggcctgcttt ctctccagggcctggctctg 33540 cctcgcttct ctgtattaac ccatgtcttg gtgcttcttt ctcttggggattccttccga 33600 cacccccagg ctttggggtg ctcctgatcc catgaatgcc tggttgcccggggtctctgc 33660 ctgcccagga accccgagga accagtctct ggctagctgc cggccctcgcagcccagagc 33720 tgtccttggg ggagccaaga gtggcagtgc tgccctgacg gtgtgagaggcagccctcta 33780 tgcagaaggg ccctagccag gtctgcgtgc ctgtgcgtgc atgtgtgcgtgtgcgtgcgc 33840 atgcgtgcgt gtatgtgcat gcatgtgtgt gcatgtgtgt gcgtgtgtgcgtgcgtgtgc 33900 atgtgtgcgt atgggtgtgt gcatgcgtgt gtgtgtgtgc gcgtgtgcgtgcgcgtgtgc 33960 gtgcatgtgt gcgtgtgcat gcgtgtgtgt gcatgtgtgt gtgtgcgcgtgcgtgcgcgt 34020 atgctgcact aacctgccgc ttgctgactg aggtttttgt ctgtacacaggcgagtgagc 34080 tcagggggcc acctgcgctc cccccgctac cctccgagcc gcgcccctgtctcaggcacc 34140 tctcggacct cgctgtgttt cactgcctcc tgcccacaga cccaggcctgccggcccgga 34200 cccgtcccag cctcccctcc ccaccccatg cagcccccag ggggatagcccatgggcccc 34260 tgtggaccct ccctccccaa gtggacacat ggctgtgcag gccaggaggcccacagatgg 34320 actgagtgct gggaaggggc ggctgcgagg ggtatcaacc ccccgagtctctccctgaag 34380 gggagcaccg ggcgagtgca tgtgctactg ctgctacagg cctgtctatctgtttgtctg 34440 tctgtgtgtc tgtgacagtc agggaaggat gcctcggagc tgaggtggggtgagacagag 34500 tgggagagat tacggcatgg catggagggg cccaaggagc aggggctgttgacaaaggcc 34560 ttaccaggaa gggttaggac actgaccatt ctagaaatgg gtttcgaatggcacaacact 34620 ttctatttca caaaagacca aaagccagag gccccaggct ctgtgctgatgaacagcctg 34680 gctgagccct ggccctggca ggtttagggc ccatttgggg ccccctccttctctgtcagg 34740 gctggggtgc tctgtctggg aatgagggag ttaaccaagt ttggtgcaggagcaggggca 34800 gggggccact gtagtgagcg tggagaaatt tggaaacacc tatttcttaactcaaataaa 34860 gtccagtttg tacctatctg gtgtgttgtg tttttttttt cccccgccgatgtctctgcc 34920 accacgtggc cctctcagtt tcccctccct cagttccctc ttgcctccatgaatcaccct 34980 ccctggccca gcttggattg ccatctcaaa gccaggtctg ggcatgccttgctgtcctgg 35040 ccaggtgggt gggctttccc catctccaga gaacaaaatg catctctctctctgtgtctg 35100 tctgtctctc tgtgcgtgcg tatgtgtgtc tccctcaccc tgtgtgtctctgctctgtgc 35160 gtggcccccg tggctgcttt cccctcagca caccctgaaa gccggtccctggccgtgctg 35220 gcccccctgc aggacgtgga cgtgggggcc ggggagatgg cgctgtttgagtgcctggtg 35280 gcggggccca ctgacgtgga ggtggattgg ctgtgccgtg gccgcctgctgcagcctgca 35340 ctgctcaaat gcaagatgca tttcgatggc cgcaaatgca agctgctacttacatctgta 35400 catgaggacg acagtggcgt ctacacctgc aagctcagca cggccaaaggtaactcccca 35460 ctcaggcatt gggctgccgt gggtgcccaa gagctggagg gaggggactgggggtgtaca 35520 gtaagatgcc tgggaaacag agctccaacc ccgaggggaa gcgggggagggtgggagcta 35580 gtacattgcc ttggcctcaa taaaataagc acttagaata gtgcatagcacaaaggaaag 35640 gccgcaacag ggttaactgt tcctaggagg gagtgcatct gctgaggtgaacgtgggttc 35700 ccacgcctgc cctgcaagtc accagccata taactttgaa caagtcacttcatctctctg 35760 agctttagcc tgttcatctg tagaacaggg atggtgatca ttcctcctctgtagagggat 35820 tggcaggatt aatgagatag tttatgtgaa gaacaaagca cagggcctgtcaaagggcct 35880 ttccaggaga gggtagggca ctgagcattc cagaaatggg tttcaaatggcacacctgcc 35940 taataaatgc cagccattgt taccactgat gctatctctg acctgggcctgcccacatgg 36000 aagggcagag attatggcac ctgccttgct acgttgtggg tgatgaagacctaagcggaa 36060 gctggagggg ctttagaagc aggagggtgc atcgggtcaa cataagggacccttatctcc 36120 tccagaagct tctttgaggg ttggtaggtg gggccaaggg gcatctgctctgggtctggg 36180 gaaggtggct aggagcatgg gcatgacccc agcacagagg agaattctgagaagtagatg 36240 gaggaggggt gggcttggct tctaggaccc tatcagagct gggctgtgcttcatccaggg 36300 tgggcagggt gaggggagga ggggggagct ggggccaggc cctgggctcaggccctgggc 36360 tcctatggag tccccagccc accatggcag tctgcaccca cccttgctgacctccaccct 36420 ctcgaggtct agtctctgga tctgtgccca cttcctcccc tgagtacccagagcctttgc 36480 tgggctctgc ccaggctcca gcttcctgct cagccttagg tcaggagtggtgggttggga 36540 tgcctgggcc tccttagcct tccctatctc tgagctgccc cctgccccacagatgagctg 36600 acctgcagtg cccggctgac cgtgcggccc tcgttggcac ccctgttcacacggctgctg 36660 gaagatgtgg aggtgttgga gggccgagct gcccgtttcg actgcaagatcagtggcacc 36720 ccgccccctg ttgttacctg gactcatttt ggtacggccc ctgtgctgcaggtgtttgag 36780 ggccccccca agggcccagg cggcgatggg gtgcacccag agggcagggcccctcactgt 36840 gcctgctctg cattcccacc cctcctttct gcaggctgcc ccatggaggagagtgagaac 36900 ttgcggctgc ggcaggacgg gggtctgcac tcactgcaca ttgcccatgtgggcagcgag 36960 gacgaggggc tctatgcggt cagtgctgtt aacacccatg gccaggcccactgctcagcc 37020 cagctgtatg tagaagagcc ccggacagcc gcctcaggcc ccaggtaccaccggggcccc 37080 aaatgatgct ggggctgcct gtgaggggcc agcccagccc tggggtgggaggcacggccc 37140 tgggcctgtg ggcagctgtg tggtcttgca gctcgaagct ggagaagatgccatccattc 37200 ccgaggagcc agagcagggt gagctggagc ggctgtccat tcccgacttcctgcggccac 37260 tgcaggacct ggaggtggga ctggccaagg aggccatgct agagtgccaggtgaccggcc 37320 tgccctaccc caccatcagc tggttccaca atggccaccg catccagagcagcgacgacc 37380 ggcgcatgac acagtgtacg tgtctgggaa gttccccggg agtgtcccctgcagcaccca 37440 cttggcttgc aatgccctgc ccctctcccc agctctcccc aggcctttcctctgtagcct 37500 gaccagggac agggtgcctg ggggaaggga acccggaggg actgtgaggtcattgcctcc 37560 cctgcaagcc cacacagcac tcatctgctg agtcacccct cagtgcccgttagcactgac 37620 tgggcaggca gtactgcctg ctactaaccc gcatcgggcc catgagcaagttacagaagc 37680 cctctgtgcc tcagtttctc atctgtaatt gggttgttac gagactaaatcagttaatgc 37740 atataagccc cagagcaggg cctggcacct ggcaagcagc tgggaggtgtgaagtctcag 37800 tattcttgtt ttagtagcca ttatcatcag cggtggtact tcctggagacattgcaatga 37860 aaaagcaggt gtgggcagtg tctggcatgg aagggatgct ctgtccagggttgctaacaa 37920 atagcaaata acgaagaagg ggccagggga gacacagagg aacgtatttgggttggtgtc 37980 ctcttagcct gggatacttt cttatgggag catctcagtt ccatccttgaaggaatctga 38040 gtcttgtgtg agaaggtctg cccccctccc ataagacagt gcccactctttgacccacga 38100 tgatttcctc ttatggctgc aacatgacaa agatcacttt cattaagtgctttctaagtg 38160 ccaggtcctg agagctaaag gtatgacaag gaccatctta ccttgccacaggcctataag 38220 gcggatacta ttagccccat ttacagatgg ggaaactgag gcttagagagatacaggaag 38280 ctgccaagag gcaagaaagg actgtctgac cctggtgccc agctcttaaccaggatgtcc 38340 ctgtcaccca tgctgcatcc tccctgccca cccgcctgcc catctccagtgatgccaccc 38400 cggctccaac ccctgcccac agcctcccat gactgtcatt ctcagcagcaggcctccctg 38460 tctcccagtc tgctctgagg ccctgggtga gtgtctgtct caggatctaagctggcgtct 38520 ccgcttctgc ctgtatctgg ggtcactggc agcccttggc ttctcttctcttataaatag 38580 tgtcagcaga gataaatgaa tgggtgactg ttctatgcag agataactgcaaagagaagg 38640 gaagggtgtc tagggacagc cctgacatga aggaggacag tgcaggcctcctctctgtgt 38700 cttcgccaga gacagcctcc ttatcatcca gggagcaggg agtagggaaagagctttgga 38760 atcacatggt accaaggtca aactatcatt tattagccgt gggaccttgaacaagtcaat 38820 atctctgacc tcattggtaa aatggggata attttgtaga gttgttgcaaggattcacga 38880 gggggaaagc atgtcaggta cctggtatag gctgggcaca gagcaggcaactcttgtaat 38940 atatcataga atgtatttgg tacttactgt gtgtgaggtt ctgggtctggtgggacaagc 39000 agatctgaca acgtcagccc tctctttaag gagcttgcag accaggtcaggagctatgac 39060 tgatgcaaga ggaagagccc ccgcccatga aaagcagcgt gggccagtgcccatcgatgt 39120 gcaggcgaga gggttggggt cctgggagga gtcagaggat gaggggtagagagcgggcca 39180 ggggcagggg ggcttcatgg gctgtaccca cttaaactat gtcttgaaggacaaggaagc 39240 cttaggtaag aggagagcat tccccaaggg aggaatgttg tgagcaatgacctgcagatg 39300 gaactcggtg tctcgttcag gggcctgata gttagccagc atcaagatgcagaggagcaa 39360 cagagacgtg gctggcaggg agtttgggtt gagggctttg ctgtcctcctggagggcttc 39420 cctgcagctg tgctggagac agaatggcag tggccttggg gatggagaagaagggcttag 39480 gggaggggcc ttgggaacaa agctttgttc gagttttgac atagaggttgtgaccatgca 39540 gaggtgggaa gacgaggggc cagtcaacag aggcagggac cccagagggtggcccagtgt 39600 aggaggaaga gaagatgata agttgcctgg cctgacacac tggagggacagaccagaggg 39660 gccagggcag atgtagactt ggaagccagc ccgggcccag acccagactttgctgcacca 39720 tggatgcact tcttgctgcc tgccccatcc ttgccccatc cttgcccactcgcctcctcc 39780 ctgcagtgga tgggggtgag ggaccaggcc cgggatggca tgggcctacccctcaaggta 39840 tcctccgggc tcaggcccag tgtcactgtc cctcccctcc cagacagggatgtccatcgc 39900 ttggtgttcc ctgccgtggg gcctcagcac gccggtgtct acaagagcgtcattgccaac 39960 aagctgggca aagctgcctg ctatgcccac ctgtatgtca caggtgaggcaggcaccctc 40020 gtggtcagct gcacgcacag cctggcctct ggcactacgt gggggctcagggaaaggggc 40080 ctccacccag ctcccttccc ctccatcccc tggggaccct cttgccttgcccctgcccct 40140 gcggctgagc ccccaggccc tagcctcctg ccctgaggct cggtgatcctgtggggctgt 40200 tgggcccttg gacccagcag acattcgaac tgcggctttc agatgtggtcccaggccctc 40260 cagatggcgc cccgcaggtg gtggctgtga cggggaggat ggtcacactcacatggaacc 40320 cccccaggag tctggacatg gccatcggtg ggtcagggct gcacagggccatgggtgggg 40380 aaggggtgtg gagaaggcag gctcaggcag gacaccatgg gggccaggccccagaagcgg 40440 atgggcaggg gcaggagctg atggaatgct ggtgggacca gctttgcccgtcttctctcc 40500 acgttgcatg gggctcttgc tctgggtgag gagaggaggc acggggcactgccacattcc 40560 cttcccatcc tcagagtggg tgcctgggtc aggacttgca aatgccctctcctcgtcttg 40620 tccaggatct cctcccgctc tgcctcagtt tccctacctc aggtatcaaggaattagagt 40680 tcaattccag ccccatctgt gcctgtactg gtaccttgtt ggctcttaacctcccaggga 40740 agtggctggg caagagcaga tggggggaca ggcaggaagc aacagcagagactgaggcac 40800 gtcatcagag cagactaatg atgagttcca gggtcccggg ccagctggatggggaggggt 40860 tactgctcct gcaacagcag cctctagtag ctcctctccc gccagacccggactccctga 40920 cgtacacagt gcagcaccag gtgctgggct cggaccagtg gacggcactggtcacaggcc 40980 tgcgggagcc agggtgggca gccacagggc tgcgtaaggg ggtccagcacatcttccggg 41040 tcctcagcac cactgtcaag agcagcagca agccctcacc cccttctgagcctgtgcagc 41100 tgctggagca cggtgagcct gggtgctcct gtcgggtggg ggtgggagctgctgggatgg 41160 ggaatggggg ccctgtggtg gaggctcaag ggatggcctg gacatggtaactggcaggag 41220 cagcctagcc gggcgggacc ttggcccatc tgtacacttc cttctccctcctgaaagcag 41280 cagggcacgg tggctgaagc tcaggctttg ggatcgggcc tgctggggtccaaccccaca 41340 acctcagctt tgctgctctc tggctgtgtt cccctgacaa atcgctaaacctctctgagc 41400 ttcagctttc ccatctgtaa aaacggaact caagtgttga tgaggggtgttagaggagtg 41460 gtgggtgctg aggacctgat gtcaagccca gcacagagcc tgcgctctcctcctcccagg 41520 cccaaccctg gaggaggccc ctgccatgct ggacaaacca gacatcgtgtatgtggtgga 41580 gggacagcct gccagcgtca ccgtcacatt caaccatgtg gaggcccaggtcgtctggag 41640 gaggtgggcc cctttcccac atgtggcagc ccaggtctgg cccagcctggccggaatgcc 41700 ctggggcaag atctgggtga cctccctgtc atgtgtcccc tagctgccgaggggccctcc 41760 tagaggcacg ggccggtgtg tacgagctga gccagccaga tgatgaccagtactgtcttc 41820 ggatctgccg ggtgagccgc cgggacatgg gggccctcac ctgcaccgcccgaaaccgtc 41880 acggcacaca gacctgctcg gtcacattgg agctggcagg tgggtgacagcgggccttct 41940 tcctagcctc cctccaaggc ccaaagctct ctactcacac ccccaggtacacaacctgcc 42000 tgacactgct gcagatccaa acccatgtcc tctggtcagg cctgtccatgtcatggctat 42060 acaaatacca tattagtaat aatgacaaca atcatactaa caagatttattggccagacg 42120 cggtggctca cacctgtaat ctcagcactt tgggaggccg acacaggtggattacctgag 42180 gtcaggagtt cgtgaccagc ctcgccaaca tggtgaaacc ccatctctactaaaaataca 42240 aaaattagct gggtgtggcg gcaggtgcct gtagtcccag ctacttgggaggctgaggca 42300 ggagaatcac ttgaacctgg gaggcagagg ttgcagtgag ccgagattgctccattgcac 42360 gctagcctgg gcaacaagag caaaactctg tctcaaaaaa aaaaaaaaaaaaagatgtat 42420 tgagcctagt atgtgccagt ctcagtacta agcactttac atgtattgcctcatttaatg 42480 tttacatcag ccttgtgagt tggggttggt tattatcccc attttacagatgaggagact 42540 gaggccgagg ctaagagtaa ctggcccaag ttcacagaac ctaataagtacaggagctgg 42600 gttcaagtgt ggctgcctga ctcctagctc ctgtgttaaa cataataggaaatagtatct 42660 cagatataac aaacatggga agaccaagtt ggtttttaaa gaaacccatgggcacatctt 42720 atcaccgaac tgccagccct gagaatcctg gccgcccttc ggcacaagcctgtttgacag 42780 agcctcccaa tgtttgcagc agcaaggatt cttttttttt tttttttttttgagacagtc 42840 atctcactct gtcacccaca ccatctcggc tcactgcaac ctccacctcccaggttcgag 42900 tgattcttgt gcctcaggct gccaagtagc tgggactaca ggcttgcacaaccacgccca 42960 gcaaattttt tgtatttttt agtagagatg gggttttgct atgttggccaggctggtctg 43020 gaactcctgg cctcaagtaa tccgtccacc tcggcttccc aaagtgctgagattatagat 43080 gtgagccacc gcctcgggcc ttaggattct ttttataatt tttcctttaaagatgtagtt 43140 tctcaaacta gttactacct aatgcattag gtagtaactg tttggttttctaatttgtta 43200 attaactcta gacatatgta accgccactc agaactcctg atcaacatgggctaaacagg 43260 gttacctgct gagtaaatac aattccaacc aaaagcaaag aaagtgacattttacttagc 43320 acaggcaatc ttatcatggg taaatggaca ttttctggtg ctttcaaaaaaccattgaag 43380 ctcacttgaa actttccggt gctttgacct tcatagactg cccccaccacctcccagccc 43440 atgggggaca ggattcctag aggcgagggg aggcgtctgt gaagtggaagataagtggca 43500 atagggtgtg taacaaagag acagggaacg cttctggccg tgctgtgaagatggtggggg 43560 tgctgggcgc tcttttaagt gtctccctcc ttgcctgccc tgtccctagcactctccact 43620 cagtcaccac tttactcctt gacagtccat cttcaggact cacctttcttcccagatata 43680 ggaggagccc tgaagtttgg ggagcccctg gccctgagag ccctgcagcacagcctagct 43740 ggaacagtga ccgatggagc cccatttgag cccagccagt cagggctctgatggggctgg 43800 ggcttgcact gacaccccta ttccactaga ccccacccag ccacacccagacagggcaag 43860 tggagtggtg gccccccttc ctccacgtca acctcactgc agtgtcttccccacccccgg 43920 gatggcctcc cccgttgtct gtgtgcctgt gaagggtggg ggtccacacccggccgccct 43980 gccttcccaa gtgcctgact catcctactt ctcctcagtc tgagatgctcaaacacgctg 44040 gggtgggggg tgggggctgg cgggggacac ctggaagtca caggctcctttgcaaagcac 44100 attgcaatgt gtatttaatt ccatcatgag aattagaagc tgctttaccaggaaatcata 44160 tgaccatgtg atattatgga gaatgaaatc acaaccacat gtgggagatactgagagtcc 44220 atttctagat ggtttggtta gtatttgggg gttacttgtt tgttcatcttgccatctccc 44280 agatgctatt aatagaagcc cctctaaaca tttttcaaag attcagtgaaaaaacttcaa 44340 ggtagaggca atgccatcca tgagcttaat tcagggtcac aggctgagccacctgcaggg 44400 gccaagttca ggtagtcaga agccagcggg tggccctgtg aagaagggcggccacccaca 44460 gggggcagca ttgggccact cctctcccgc ggattcatgc agcgaggggagcgactcagg 44520 atggccagtc ctgatttttc aagagaggca ggaaatccag atttcttttgtgtgtgaaaa 44580 tctcctaatt tcaaagctgc caaattaata cagaaggtca aacaaagcaagcctgcacct 44640 tgtgccaggt ttcctcagat ttggagaaca tcacgtttgg atttacggggcggggcgggg 44700 cttggtccag gcagggaact ctgttgtccc agttggtggg ggtggggggtggtggcttgc 44760 agggaggcgg gggtctagcg ttctgactga gccagtcact gatgggggatcttgggaggg 44820 gctgaggagg cagggtaaag ccagccccta gccccttctc ttcccacccctatccccatg 44880 tgtttcagag gcccctcggt ttgagtccat catggaggac gtggaggtgggggctgggga 44940 aactgctcgc tttgcggtgg tggtcgaggg aaaaccactg ccggacatcatgtggtacaa 45000 ggtcagagtg tgctgctggc tgagcctggg ggagggagga ggggctccctgggggcgtgg 45060 gagggtcctg gaaggcctta ggagggcgga gcccgggcag aggcgtggttaggaggagga 45120 aaggggctgc agaggactga ctagctgagg ggtgcagggc tttctgtgggagataaggga 45180 ggagctgact ctgggtcctg gtgagagatg cgctgcccag agtaggagatgaggccctgg 45240 ccccaaggta gagatgaggc caggcccagg ctgaaggtga gaccccactctgcaggacga 45300 ggtgctgctg accgagagca gccatgtgag cttcgtgtac gaggagaatgagtgctccct 45360 ggtggtgctc agcacggggg cccaggatgg aggcgtctac acctgcaccgcccagaacct 45420 ggcgggtgag gtctcctgca aagcagagtt ggctgtgcat tcaggtaggcaggagttccg 45480 gagaaaggta aagcgcacac cccctggaat ctgatgtgac cctccatgctctgcccagga 45540 agcagccagc cctggactcg agccaccaca cccccagccc agcaccccgccttgagcccc 45600 caacattctt gcacctcttc tctcttcttc ttctgtccac ctgtcccagtctctggcctg 45660 cttgctttct tcccctccca cctaacacca tgacatctct gccccagctcagacagctat 45720 ggaggtcgag ggggtcgggg aggatgagga ccatcgagga aggagactcagcgactttta 45780 tgacatccac caggagatcg gcaggtgtgg ggctaggagg gaagccagtgggggccgaga 45840 gaggctgctg ggtctgaggg ttgggagggg tggagagggc cacagtgatggctgatctct 45900 gaccccctcc ctgtgtcaac caggggtgct ttctcctact tgcggcgcatagtggagcgt 45960 agctccggcc tggagtttgc ggccaagttc atccccagcc aggccaagccaaaggcatca 46020 gcgcgtcggg aggcccggct gctggccagg ctccagcacg actgtgtcctctacttccat 46080 gaggccttcg agaggcgccg gggactggtc attgtcaccg agctgtatcctgggacaggc 46140 tgggggctag ggggatccat gcctaaatga ctggtccttg taacatccaataggcaacat 46200 gttttacagt ttacaaagta gtcccaattg acaattggga gggatacatctggagacttc 46260 tatgaaaacc aacagtttta agcccattgt tagcttaaat tttcatagcatttaatgtgt 46320 gtgcctggca gtgttctaag agttttacaa atattaacct atttaatcccatatattcct 46380 atctcctgga attttattat taacttgtat tatccccatt ttacagatgaagagacaggc 46440 atggagaggt taaataactt gcccaaggtc acagagctag cacatggcctatgcttttca 46500 tcatgaaatg cagtacaatg catctgcata tttgtagtga ccccatttcagatcaatact 46560 tggctttatt tagaggctgt tttcataagg aacagatttt accctttcatgaaggctgtg 46620 gttaagcaaa ggcacattga ggtaaggatg ctgggtgatt ggctcatcaccccagtagca 46680 gggtgaccgt aacctgcaca tgagcctcct gcctgcactg tttgaattcaaacctcagca 46740 aaatgacaaa ttattccatg tcatggatct tctaggtcat tctcaaatagagtcaagaag 46800 ttccatccac agtcaacaaa attctgctgt aatttgatcc ctataaatgctttacagggt 46860 gggcaggaaa ggaaatgtta ttcccatatt actcctggga gaacctaggctcagagaagt 46920 gaagtgactt gtctgagatc ccatagaaag tgggagttca agcagggatgacccacctgt 46980 gccctcagag gacaggaggt ggggaggggg tacacgtgga ggagcggagaggcagtctct 47040 ggctagtatc aagcattctg taaggggaag gagaaccccg tgctgagctgggacctgccc 47100 tgagcgctgg gctgggccgg gcagttggca ctgggcactg ttctccttgatctgggatgt 47160 agctgcacag aggagctgct ggagcgaatc gccaggaaac ccaccgtgtgtgagtctgag 47220 gtgagggcag tgggtggcag gggccaggtt gggcaccagc cttcacccacctgagctttg 47280 agaaccaaca aatgggtcct gagtgtgcca tctgtgccca caggaagccaggggtggagg 47340 tggagagcac tgttaggtag gcagcagagt ccccacccaa tgataccagacccccatcta 47400 tctgagactt gtgctattct ctctaaatct cagcaaaccc accccagctcctaactgttg 47460 tttcccactc ttcatcacat aaacatcccc caaacatttc tcctggaaggagccccacct 47520 agattttatt gatctcactg cagtcctgcc acgtcaggct gtacatgttgtgcactgcac 47580 aattctactt attaccacgc aatggctcct ccctataatt gtgctgtaccctccggtgca 47640 tatggtagcc ttggatcttt gcaaccccaa acctgtttca gccccttccacgagccatct 47700 gaaggctact ccacaggcac agccggaccg cttgccgccc tggaggtgttcagacataca 47760 ccacccttcc ccctcagact ctgggcccac tatttccaca gatccgggcctatatgcggc 47820 aggtgctaga gggaatacac tacctgcacc agagccacgt gctgcacctcgatgtcaagg 47880 tgaggtgggg actggagagc agacagcccc tgtgggagcc aggaggtgaagcatccttcc 47940 ttgttcattt ggcccgcaca cctcgccttg tgtcttccag cctgagaacctgctggtgtg 48000 ggatggtgct gcgggcgagc agcaggtgcg gatctgtgac tttgggaatgcccaggagct 48060 gactccagga gagccccagt actgccagta tggcacacct gagtttgtagcacccgagat 48120 tgtcaatcag agccccgtgt ctggagtcac tgacatctgg taaggctggcatgctgggct 48180 gggccgacca gggcagctgc ccttggggct gtgctgggga cgcgctcactggcagggaga 48240 tttaccgagc ctgaattcct cctgaaggtg ggctggaggc attgtttgcagggtctcctg 48300 cccatgttac tccttgcccc ttgtgagtca gggctgcccc attctctcaaggcctcagcc 48360 ctgttagtcc ttgactcctt gtctcccctg gaagccgggc cttcccctcagcattcagcc 48420 tgcctcctcc agtaaggcag gcatggtccc attggttccc aggcttccctgggcttcctg 48480 ggccagccct gccatgaccc tggacttctc caggcatatc tggacctgtaggttcagggt 48540 cctccctgaa gaagccactc ctgtgcccat tgtccatggc agtgttcccagggaggtaac 48600 agctcactca ggtcagcagt agcaaagaac tgctcccttc cagtcagagaggggcggtcc 48660 tcttacctat cactctcctt ttcccacagg cctgtgggtg ttgttgccttcctctggtaa 48720 ggacccctct gcaatgtccc agcagtctcc tggcaggtct acccctaacctttgcagggc 48780 tgcagcccac ccccttctct tccgcacccc ccactccttc ttgcactgcaaggagcctca 48840 tgtgcatgaa ggtggacacc cctgtctgca tgcccacact ctgcctgtccccacacccct 48900 ccataagagg tgggcaccct agatggagag agcccagcgc aggctcagggccatggaggc 48960 agggaactcc ttggctctga gtgtccaaaa cttggactag atgggagtggagctcagggt 49020 gggcacatcc ccttggcaca gactcttcac tcatggagag gccacactggaggagggatg 49080 gaacaggtcc tctaaagcac aggccactag gccccaaggc agcaccacctccctgcccat 49140 caggggggct ggggagggga cagggcagga gagagtccgc agcctcacctcttacccaca 49200 tttgcccagc ctctgtcatc ctcacaaccc cagagcctcc atctgtccccagccctgtgc 49260 ccccactgac attccccttt gtccccgcct gcccctcatg acagccctcttcacccctgc 49320 agtctgacag gaatctcccc gtttgttggg gaaaatgacc ggacaacattgatgaacatc 49380 cgaaactaca acgtggcctt cgaggagacc acattcctga gcctgagcagggaggcccgg 49440 ggcttcctca tcaaagtgtt ggtgcaggac cggctgtgag tacaaggccctgggagcccc 49500 cacctgcagg gtcaccctca taccacctgc ctgctactcc caaactcctgcccctcgaca 49560 tgcaagcccc caactcctta ggagccctgt ttgctcagtt attgactcactgatgactga 49620 acgataaatc ccttcttaat cctcattcat tcacaggaga cctaccgcagaagagaccct 49680 agaacatcct tggttcaaag tgagtctagt ctgcaaagtg gtggcacaaaaggtggaggg 49740 agagagaatg ttactaacag ctctatttat tgagtaccta ctgtgtgcagtaaccacgtt 49800 aggcattgta tgtacatatg acgtattagc ctcacaatag ctttgcaaaggaaggcatta 49860 ttagtcccat tttgttgaca aggaaacagg ctttaatcag tgatggagttgggggtatac 49920 atactggact ccagggcata cgtctggacc tctccatcct gggcgtcctcagtggagttc 49980 tcccccatgc ttgagaccag accctggtct tcctgacttc tgtctgtccatctctgtcct 50040 gcactggtcc cacacaatag cgttgggggt gaccaggcag tctcagccccttggttatga 50100 gcttcatgtg ggcaggttct cggttctcac tcattcatct tcaaaccccagtgcctcagg 50160 gcacagtgcc aggcattgat ggggtcttgg ggatttggga gggggggttcagcaaatgag 50220 tctggaaaga gcgcctgaat aaactgttca tggagggttg tgccaggtcacttgaaggct 50280 gaggtcattc gggtatcagg agttgaatta agagccttct ttctcagaccggaaatgagc 50340 tccagaagag agagcctggg tgggtagctg agggaagcat ccgtcttggtctggaccacc 50400 aaggctccag atgtctgggg tgttggccct acatggagac agaggggagctggggagcca 50460 ggacccgggt aaagggccta agatcacagg cctcccaggg cagctggtttaccaggacag 50520 ggccatgagc cctggtggaa gctcagaagc ctacctaggg gagccaccagtttcctgcct 50580 ctcccctggg gggttcaggg gatttgtctt taggtgtgca tcttggctgtaggcattgtc 50640 ctgacagacc cagggggaag gggaccccca ggagcccaga ctcagtgctgctcgatccac 50700 tctctcctgc caccccgccc catggacttt gtctctctgt tgccaaacctggagggtcta 50760 ggttgacagc tttccctcaa gccctctttc ctgggtttgc agactcaggcaaagggcgca 50820 gaggtgagca cggatcacct gaagctattc ctctcccggc ggaggtggcaggtaagtgtg 50880 gcaggccagc ctctgtgctt tccaccttct ccttttctct agcactgccttccccctccc 50940 gtgggccttc atctcctgct cctgtcttct cgctttcact ggctccatgcctagcttcct 51000 gcctgttccc tgaccctctg catgctcagg cctcttcccc agggctgaggtgggcctggg 51060 ggggacaatc ctgccccagg ggtccctcag gtctgactcc agtaccctgtctccagcgct 51120 cccagatcag ctacaaatgc cacctggtgc tgcgccccat ccccgagctgctgcgggccc 51180 ccccagagcg ggtgtgggtg accatgccca gaaggccacc ccccagtggggggctctcat 51240 cctcctcgga ttctgaagag gaagagctgg aagagctgcc ctcagtgccccgcccactgc 51300 agcccgagtt ctctggctcc cgggtgtccc tcacagacat tcccactgaggatgaggccc 51360 tggggacccc agagactggg gctgccaccc ccatggactg gcaggagcagggaagggctc 51420 cctctcagga ccaggaggct cccagcccag aggccctccc ctccccaggccaggagcccg 51480 cagctggggc tagccccagg cggggagagc tccgcagggg cagctcggctgagagcgccc 51540 tgccccgggc cgggccgcgg gagctgggcc ggggcctgca caaggcggcgtctgtggagc 51600 tgccgcagcg ccggagcccc agcccgggag ccacccgcct ggcccggggaggcctgggtg 51660 agggcgagta tgcccagagg ctgcaggccc tgcgccagcg gctgctgcggggaggccccg 51720 aggatggcaa ggtcagcggc ctcaggggtc ccctgctgga gagcctggggggccgtgctc 51780 gggacccccg gatggcacga gctgcctcca gcgaggcagc gccccaccaccagcccccac 51840 tcgagaaccg gggcctgcaa aagagcagca gcttctccca gggtgaggcggagccccggg 51900 gccggcaccg ccgagcgggg gcgcccctcg agatccccgt ggccaggcttggggcccgta 51960 ggctacagga gtctccttcc ctgtctgccc tcagcgaggc ccagccatccagccctgcac 52020 ggcccagcgc ccccaaaccc agtaccccta agtctgcaga accttctgccaccacaccta 52080 gtgatgctcc gcagcccccc gcaccccagc ctgcccaaga caaggctccagagcccaggc 52140 cagaaccagt ccgagcctcc aagcctgcac caccccccca ggccctgcaaaccctagcgc 52200 tgcccctcac accctatgct cagatcattc agtccctcca gctgtcaggccacgcccagg 52260 gcccctcgca gggccctgcc gcgccgcctt cagagcccaa gccccacgctgctgtctttg 52320 ccagggtggc ctccccacct ccgggagccc ccgagaagcg cgtgccctcagccgggggtc 52380 ccccggtgct agccgagaaa gcccgagttc ccacggtgcc ccccaggccaggcagcagtc 52440 tcagtagcag catcgaaaac ttggagtcgg aggccgtgtt cgaggccaagttcaagcgca 52500 gccgcgagtc gcccctgtcg ctggggctgc ggctgctgag ccgttcgcgctcggaggagc 52560 gcggcccctt ccgtggggcc gaggaggagg atggcatata ccggcccagcccggcgggga 52620 ccccgctgga gctggtgcga cggcctgagc gctcacgctc ggtgcaggacctcagggctg 52680 tcggagagcc tggcctcgtc cgccgcctct cgctgtcact gtcccagcggctgcggcgga 52740 cccctcccgc gcagcgccac ccggcctggg aggcccgcgg cggggacggagagagctcgg 52800 agggcgggag ctcggcgcgg ggctccccgg tgctggcgat gcgcaggcggctgagcttca 52860 ccctggagcg gctgtccagc cgattgcagc gcagtggcag cagcgaggactcggggggcg 52920 cgtcgggccg cagcacgccg ctgttcggac ggcttcgcag ggccacgtccgagggcgaga 52980 gtctgcggcg ccttggcctt ccgcacaacc agttggccgc ccaggccggcgccaccacgc 53040 cttccgccga gtccctgggc tccgaggcca gcgccacgtc gggctcctcaggtgaggagg 53100 ggcaggggta gggcagcagg tgcagaggag ggtggggtgc gctggagagaggctgtggga 53160 ggagcagagg gctggggaca cccaccaggg gcaggctgag gccccgagggtggaatcagc 53220 agggctggag gggaggaaag caggaatggc ggcagggctg ggtgggctaggggttccttc 53280 tggttctctg ggctgagggc tgcagagagg tgggaacttg ctggtactgactgaacaaat 53340 actcacgggc ctgagtcttc acagccccag gggaaagccg aagccggctccgctggggct 53400 tctctcggcc gcggaaggac aaggggttat cgccaccaaa cctctctgccagcgtccagg 53460 aggagttggg tcaccagtac gtgcgcagtg agtcaggtaa taagaggcctgctgggtgag 53520 gaccctcctc ccctcctgcc ctcccctacc cccatcaggg agcagtcatggctggtgaga 53580 ggtgggccac cttgacaaac ctagtggaag gggtctgctc agacaactataacaatagca 53640 gtagctgaca ttcattagat aagctgagtg ttctattaaa cactttacaagcactgcctc 53700 attcaatcct gcagcactgt ttgggaaata ttagtatcat tgtctctattgtacaggtga 53760 ggaaacgggc ttagtgatgc taaggatctg tccaagtcgc agggctagtaagtggagcag 53820 ctgaagttgg actgtgtgac cttctgcagc caggtccctg aaacggcttcaggacaccac 53880 ttatgtctgt cgggctggcc tcctctctcc tgggagaccc tagaatgtttctgtaactgg 53940 ctgtactttt caaggagctc aagatatagg gccctcctgc ctccacagtccccctttaga 54000 tgtgtgtgtg cttgggtgtg tacccaaaga cactcacttt ctctccagcctagaggacca 54060 cgacctggat tgtgccccct aagtctccat tgctctgcag ctccgaacacctgactgccc 54120 ctccctgacc cttctgcaca gaaaagcagc ctttggagct ctttgctagctctttgccct 54180 cttctgtttc tctgcctgag tgtcctggag ctccagatag ggaggcattccccatgtggg 54240 gtcaccccat cccccctgaa aaaggggcat tactcaagga cgacaagcaaaggcttcggg 54300 aggttgggtt ttccaggcca gcatgcagga agggagcgag tccatgaagccaggctgtgt 54360 gcaggatctt gacaagccac cagtcctgtt cttcccccat ttcctggtaaaactcagaat 54420 agagtggctg tcgagtcttg cctcagctgg gcttataggg attgtgtccagccttggcca 54480 gggcaaaggg ggctgcagtg agagaaaagg atggagggaa gggctgctggtggggagggg 54540 agggtggaga gtgaggggga aagaaagcga tcagccagca gagaggcctggggacagctg 54600 atcccttccc acctggggcc tccctcccgg cccagttttg cttatgcagctgatttcctg 54660 ctgaggcagt gtccccttac ctcatccgct ccccgttccc tgcagaaacaaggctgtccc 54720 aggctacttt aacaaccagg gctggcacct aggaatgggg gtggggctggcggggctggc 54780 agggctgggc ccgggtcact ttcacctctg agagaggtgg cctctctctctctctccctg 54840 ctacccagta ccctcacttg gcctggaggc agccattgag aaactgtgttcacattgcct 54900 tgttggaacc tcagtgtggg acccctcctt ggggagcagt ggggtacagcgggaaggggg 54960 cacactgcca tcctgatcac cacacctgct gagtactcct cttctgcctggctcatcccc 55020 actcccagtc ccccacaatt cttcagacag gcagcagctg gggctcacaaggctcctcag 55080 ctttctcttc ccaggtgaat aaaatccacc cccaagtcct cccctatccccacccttcac 55140 ccaccacccc catggcccag ctgggattct tctaaaggga cattcccagggatcatgcac 55200 tcaaatcctc agggcactaa agaggcacag gctggctaca ttggaggaaggaaaactggt 55260 cgcatcccta gccctgcatg tgcacagcca ccagctagct gtgaaggcagcttctctgct 55320 ttgctggaac agcctttctc tgggggtctg tctgctccag gcttctctgctggccatatt 55380 attgagaaaa tgataacaac aagagaagtt agtatttatg atcagttactctatggcaga 55440 cactttacat gcctcgtaat tgtcacagca agcatcctgt gggcgtgggattattttcct 55500 tgatttacag attagaaatg gagcttctga gagagtaaat gacttgcccaaggtcaggca 55560 gatactgtct cttgcccctt caaaagctct caccacttaa ttgacctgaaggagtataca 55620 gaagacctac gaccctgtcc cccgatggca ggtgggaaga ggggccaggaagccgacacg 55680 cttccagggt ctgcccacag ttttcctaga ctgcagctct tttgagactgcacattctga 55740 tagaacattc ctcatttggt cccatctcag ctcgggtcac taactcatctcaattctctt 55800 ctcacttgcc ctgtggtgcc ccagagaggg taggtcttgc aggatcttaaccattatagt 55860 ttttcagtat ttgtttgctt ttattttatt taacaaatgc ttatagaccacttacttagt 55920 gccaggccct gctctaagtg atttacaaac ccatgacata agtagcattgtcagcagtca 55980 gtgcaaggaa aaagcagttc tgcacacagt gagggcctgg ggaaagtgatgcttgcccca 56040 aagaaaatta tcgaaggcat ggcgtggggt tccactttcc acatctgcctgggaagagac 56100 aaagtttcat gctgcttcct gatggctgtt tgcaggcctt ctccacccctccctcagcag 56160 taagtgggcc agggtcccta cagatagatg gctgtctctg cttttcctccagacttcccc 56220 ccagtcttcc acatcaaact caaggaccag gtgctgctgg agggggaggcagccaccctg 56280 ctctgcctgc cagcggcctg ccctgcaccg cacatctcct ggatgaaaggtaaggagact 56340 ctgtctccca cagagaggga ggccagcaag tggccctgag cccaggggatgggaggggct 56400 aggccggagt ggggactgag cacggttagg ggggatgctg gagtggggagtgagtgaggg 56460 ggcctggaca tgtgctgcct cactcagcag caactcctgc tcctccctgtccccagacaa 56520 gaagtccttg aggtcagagc cctcagtgat catcgtgtcc tgcaaagatgggcggcagct 56580 gctcagcatc ccccgggcgg gcaagcggca cgccggtctc tatgagtgctcggccaccaa 56640 cgtactgggc agcatcacca gctcctgtac cgtggctgtg gcccgtgagcctggggcagg 56700 gccccagggg ggtagtgatg gggatggtgg gacagggctt gaggggttcttagctagggt 56760 ataggggctc actgggactc ttctttctct tgccaggagt cccaggaaagctagctcctc 56820 cagaggtacc ccagacctac caggacacgg cgctggtgct gtggaagccgggagacagcc 56880 gggcaccttg cacgtatacg ctggagcggc gagtggatgg tgaggatggggcagctggag 56940 ggttggggga gcggcagggg gagggtagag gagtctggta aggccagtgccctcccaggc 57000 tccacagata gcaccgtggg agctggggcc accgcttctg tgacctcagcccctccccca 57060 tactgcctat aggggagtct gtgtggcacc ctgtgagctc aggcatccccgactgttact 57120 acaacgtgac ccacctgcca gttggcgtga ctgtgaggtt ccgtgtggcctgtgccaacc 57180 gtgctgggca ggggcccttc agcaactctt ctgagaaggt ctttgtcaggggtactcaag 57240 gtcagtgcaa tggtatgggg tgggaggagg aagggggctc tgagcctagggttcttgtgg 57300 agcaccatgg ccttgcccca aggcaccacg gtgatgattt tctctctctcttagattctt 57360 cagctgtgcc atctgctgcc caccaagagg cccctgtcac ctcaaggccagccagggccc 57420 ggcctcctga ctctcctacc tcactggccc cacccctagc tcctgctgcccccacacccc 57480 cgtcagtcac tgtcagcccc tcatctcccc ccacacctcc tagccaggccttgtcctcgc 57540 tcaaggctgt gggtccacca ccccaaaccc ctccacgaag acacaggggcctgcaggctg 57600 cccggccagc ggagcccacc ctacccagta cccacgtcac cccaagtgagcccaagcctt 57660 tcgtccttga cactgggacc ccgatcccag cctccactcc tcaaggggttaaaccagtgt 57720 cttcctctac tcctgtgtat gtggtgactt cctttgtgtc tgcaccaccagcccctgagc 57780 ccccagcccc tgagccccct cctgagccta ccaaggtgac tgtgcagagcctcagcccgg 57840 ccaaggaggt ggtcagctcc cctgggagca gtccccgaag ctctcccaggcctgagggta 57900 ccactcttcg acagggtccc cctcagaaac cctacacctt cctggaggagaaagccaggc 57960 aagcagggct ggggaaggga agaggacaga ggggagtggg ccaaatgtctggagcacatg 58020 gcttcggaga gaagaccaga ctgtcctggc tggggtgggg ggaggtgctgagacctgggt 58080 tattagaatg attgcgttca aatgtgccag acactgcact gcgtgctttagccatatgat 58140 ctcatcaaat cttcacaact ctgagagaca ctgcgctatt agcatcacccatttcacagg 58200 tggcaaagct gaggttagag aagctatggg atttacctaa ggtacagagccagtgagtgg 58260 cgaaggtggg actcgaaccc tggtttctag gattgaactc tggagcccacactggaacca 58320 ctgcattctt gcccctaggg gtccctgctc tcctccgtta gccctcactatggaagtgtc 58380 cccctcctct cctctgagcc ggtggtgtcc ctccccccga cacacaggggccgctttggt 58440 gttgtgcgag cgtgccggga gaatgccacg gggcgaacgt tcgtggccaagatcgtgccc 58500 tatgctgccg agggcaagcg gcgggtcctg caggagtacg aggtgctgcggaccctgcac 58560 cacgagcgga tcatgtccct gcacgaggcc tacatcaccc ctcggtacctcgtgctcatt 58620 gctgagagct gtggcaaccg ggaactcctc tgtgggctca gtgacaggtagctgggaatt 58680 ctaggggagt agggaggaag aggtagggga ggctgggccg ggtatcatctgctccatccc 58740 tgccctccca ggttccggta ttctgaggat gacgtggcca cttacatggtgcagctgcta 58800 caaggcctgg actacctcca cggccaccac gtgctccacc tagacatcaagccagacaac 58860 ctgctgctgg cccctgacaa tgccctcaag attgtggact ttggcagtgcccagccctac 58920 aacccccagg cccttaggcc ccttggccac cgcacgggca cgctggagttcatgggtgag 58980 gggaccagct gccagccagg gtggggacag ggccctgcca gagaggcagcagccagggct 59040 caccccactt cacttacata tgtgccactt attgagtgat tactgtattcaagcaatgaa 59100 cgaagtatgt ggattgatct ttacaataac cctggagtgt ggcataatattagccccctt 59160 ttacagatga ggaaactgag gggtactgat gttagggatt tgtgcaatcagacaattata 59220 aatgctagag gcaggattca ctacagccaa aaagacagga gaatcaattattattttatt 59280 taaataagga gaaccagcta ccatcgagca ccctgctaaa tgcttgacgttcatgatctc 59340 tcttccttgg tgtggttcta caggccacac tttacggatg aggctgtggagagccaggca 59400 ggtttagtaa atcgcccagg gtcccatagc tagaaggggc aggtctgggattagagccag 59460 ccaggctgat tttgaaggct tttaatcttg gtgtcagcca cactctttgtgaatgggagc 59520 cataccttgg agccgatcca agggagcttg tcaccctgct tctcagcccctctggagttc 59580 tggggacccc gccattttgt gcctgggtta attcctcagg tgacccatatgtctcttggg 59640 gtatgccacc acctctgtcc tgcctactgg ccttcagggc ctgccactctggacattccc 59700 atggtctggt gaccaggaca ttgtcctgct gctcaagcac ccagggacctcccccgcccc 59760 cacttccctg ccaccaggaa gctgggtcag cttggcctct gtctcctgtcagctccggag 59820 atggtgaagg gagaacccat cggctctgcc acggacatct ggggagcgggtgtgctcact 59880 tacattatgt gagtgtcccc taccccaccg cagccctctc tgcccatacagtgagctccc 59940 ggactcacct tctgccaaca ccctctcccc cgtgccccac ctcccctgtacacacatcca 60000 cactgcacac tcacactcag gtgcacagta gcatggccct gagcactgtgcacctgacac 60060 taatgtcctt ctgggtctgg gtgttggcct ccggtctgca tatgtcaatcaagctatctt 60120 ccccaacagg ctcagtggac gctccccgtt ctatgagcca gacccccaggaaacggaggc 60180 tcggattgtg gggggccgct ttgatgcctt ccagctgtac cccaatacatcccagagcgc 60240 caccctcttc ttgcgaaagg ttctctctgt acatccctgg tgagtgagccccacacctgc 60300 tatcccccag tgttacctgc ccctggcctg gcctgtgcca gagatctcccagctcctccc 60360 ctgctcctag gaagaagtct gctgcttcta ctaaatggtc atactacccaccatttaaag 60420 cctgaggcag ccccgtgcaa ggcagactca ctgtccccat tccggagactggggaactga 60480 gctcttgagc tgcccaagat cacacatgta ggggtgggat ccaggactgggacatgggtc 60540 tgcgggagga cagagccccg gcagctccca gagcttcctt ccaggttcatcatccctggc 60600 tctgcctggc aggagccggc cctccctgca ggactgcctg gcccacccatggttgcagga 60660 cgcctacctg atgaagctgc gccgccagac gctcaccttc accaccaaccggctcaagga 60720 gttcctgggc gagcagcggc ggcgccgggc tgaggctgcc acccgccacaaggtgctgct 60780 gcgctcctac cctggcggcc cctagaggca cggaccacag ccaggcctcgggcttcaact 60840 ggggttccca ccaatgccac gggacattcc agggcccacg ctgagccaggcgggcctggg 60900 gcttcggtta ccaccagcag caacatctgg ctgggctctt acctcatagaccttcaagga 60960 cagagacccc agggcctgga cctgatgcca ccccaggcca aagccagagtgggagaccca 61020 ttggtcaggc tcagcagggt gggaacaggc agagggacaa gaggggaatggagaagtgga 61080 gaggaaaagg aatcgaggga caggaagggg gaggctctag gaaggttctgggttgggggt 61140 cagtgcatct cagggagaac caaggaaggt gggcatggct ggagaggaggaaaaggaagg 61200 agccccaggt gtcagggcag taggctggga gtcagtgtgg caaagcgggggcaggacaca 61260 gatacagtgg caggggccca gggctgggac atgagagaag gcagcgaggcggcagaggga 61320 gaagagagga ctcaggtgga ggtggggtgg gtcagctgtc agcatccctcagaggagaaa 61380 tgtggagagc tggaggccag cagtcactca cactcgctct gtcctcctgtccagtggata 61440 cagccctggg cgctctgctg gcccaaggat gtccccactg cccctccatggcctctggcc 61500 ttcttcccat tcatatttat ttatttattg acttttatga agtttccccttccatccgat 61560 ccctactgcc catgttgtcc tgaccatccc tcccagccat ccagctgtctgtctgtctgc 61620 cacaaggaaa taaaaatggc aagcagcata acctgtgtgt ctattgggagggatggctgg 61680 aggggaagat ggctggtgag gggtgagtcc gggacagggg catttagccctctctgggta 61740 ttccccaaca cacacattca ggaatatacc agctagcact ttgggtccttccaaccccct 61800 cccgtgaccc tcctggcccc tcacctctcc ttattcctgg agggaggggagactgtggtc 61860 tgcttctccc cttgcagttt ccggaatgtt ggcagatcca ctgaacccctgcaaccaggc 61920 tctagtagcc cccacctctt gtcacgtgtt ccctcatcac aatgtgggggatgctgggct 61980 ctgaaatagg ccagccctca ccccaatcct ggctcagcct tgttcactctccccagaaga 62040 caggcaggag ctctggtcct gacccctgga gcagagtggg tttcatcctgatggttggtg 62100 agagtaggta gtgtgaggag ctgcagaaga aaccaggaca gggaggctaaggtggctgga 62160 tcacctgagg tcaggagttc aagaccagcc tagccaacat gatgaaaccccgtctctact 62220 aaaaatacaa aaattagcca ggcgtggtgg tgcacacctg tgatctcagctactcaggag 62280 gctgaggcag gagaatcgct taaacctggg aggtgaaggt tgcaatgagccaagattgca 62340 ccactgcgct ccagcctgga tgacagagtg agactccatc tcaaaaagaaacactaggac 62400 aggctcacac cccttgccct ccatgtcaca gcacatatca gagcacatggagagcaccag 62460 ctgggagtgc tctagtctgc tgtgtccagc attttcctag ggctgagggacacaccagcc 62520 tggaccttct tgtccacatg gcaagttagg aggtctgctg ggtgctaaagcacctcaatt 62580 ctagccacac ccgtgccata gaatggtcac tgggacctag gactgagctgctctgccctg 62640 aggttgggga cgagggattg gggggttggc agggacccag acctccttgtctccagagga 62700 aatgtttccc tcatccccac cttcaaaatt ctgttcttgg caaagtaaaaggaacaaagc 62760 ctctgaccag ggtagacaga gttgtcactg ctgtgttgct gatgg 628054 3262 PRT Mus musculus 4 Met Gln Lys Ala Arg Gly Thr Arg Gly Glu AspAla Gly Thr Arg Ala 1 5 10 15 Pro Pro Ser Pro Gly Val Pro Pro Lys ArgAla Lys Val Gly Ala Gly 20 25 30 Arg Gly Val Leu Val Thr Gly Asp Gly AlaGly Ala Pro Val Phe Leu 35 40 45 Arg Pro Leu Lys Asn Ala Ala Val Cys AlaGly Ser Asp Val Arg Leu 50 55 60 Arg Val Val Val Ser Gly Thr Pro Gln ProSer Leu Ser Trp Phe Arg 65 70 75 80 Asp Gly Gln Leu Leu Pro Pro Pro AlaPro Glu Pro Ser Cys Leu Trp 85 90 95 Leu Arg Ser Cys Gly Ala Gln Asp AlaGly Val Tyr Ser Cys Ser Ala 100 105 110 Gln Asn Glu Arg Gly Gln Ala SerCys Glu Ala Val Leu Thr Val Leu 115 120 125 Glu Val Arg Asp Ser Glu ThrAla Glu Asp Asp Ile Ser Asp Val Pro 130 135 140 Gly Thr Gln Arg Leu GluLeu Arg Asp Asp Arg Ala Phe Ser Thr Pro 145 150 155 160 Thr Gly Gly SerAsp Thr Leu Val Gly Thr Ser Leu Asp Thr Pro Pro 165 170 175 Thr Ser ValThr Gly Thr Ser Glu Glu Gln Val Ser Trp Trp Gly Ser 180 185 190 Gly GlnThr Val Leu Glu Gln Glu Ala Gly Ser Gly Gly Gly Thr Arg 195 200 205 ProLeu Pro Gly Ser Pro Arg Gln Ala Gln Thr Thr Gly Ala Gly Pro 210 215 220Arg His Leu Gly Val Glu Pro Leu Val Arg Ala Ser Arg Ala Asn Leu 225 230235 240 Val Gly Ala Ser Trp Gly Ser Glu Asp Ser Leu Ser Val Ala Ser Asp245 250 255 Leu Tyr Gly Ser Ala Phe Ser Leu Tyr Arg Gly Arg Ala Leu SerIle 260 265 270 His Val Ser Ile Pro Pro Ser Gly Leu His Arg Glu Glu ProAsp Leu 275 280 285 Gln Pro Gln Pro Ala Ser Asp Ala Leu Arg Pro Arg ProAla Leu Pro 290 295 300 Pro Pro Ser Lys Ser Ala Leu Leu Pro Pro Pro SerPro Arg Val Gly 305 310 315 320 Lys Arg Ala Leu Pro Gly Pro Ser Thr GlnPro Pro Ala Thr Pro Thr 325 330 335 Ser Pro His Arg Arg Ala Gln Glu ProSer Leu Pro Glu Asp Ile Thr 340 345 350 Thr Thr Glu Glu Lys Arg Gly LysLys Pro Lys Ser Ser Gly Pro Ser 355 360 365 Leu Ala Gly Thr Val Glu SerArg Pro Gln Thr Pro Leu Ser Glu Ala 370 375 380 Ser Gly Arg Leu Ser AlaLeu Gly Arg Ser Pro Arg Leu Val Arg Ala 385 390 395 400 Gly Ser Arg IleLeu Asp Lys Leu Gln Phe Phe Glu Glu Arg Arg Arg 405 410 415 Ser Leu GluArg Ser Asp Ser Pro Pro Ala Pro Leu Arg Pro Trp Val 420 425 430 Pro LeuArg Lys Ala Arg Ser Leu Glu Gln Pro Lys Ser Glu Gly Gly 435 440 445 AlaAla Trp Gly Thr Pro Glu Ala Ser Gln Glu Glu Leu Arg Ser Pro 450 455 460Arg Gly Ser Val Ala Glu Arg Arg Arg Leu Phe Gln Gln Lys Ala Ala 465 470475 480 Ser Leu Asp Glu Arg Thr Arg Gln Arg Ser Ala Thr Ser Asp Leu Glu485 490 495 Leu Arg Phe Ala Gln Glu Leu Gly Arg Ile Arg Arg Ser Thr SerArg 500 505 510 Glu Glu Leu Val Arg Ser His Glu Ser Leu Arg Ala Thr LeuGln Arg 515 520 525 Ala Pro Ser Pro Arg Glu Pro Gly Glu Pro Pro Leu PheSer Arg Pro 530 535 540 Ser Thr Pro Lys Thr Ser Arg Ala Val Ser Pro AlaAla Thr Gln Pro 545 550 555 560 Pro Pro Pro Ser Gly Ala Gly Lys Ser GlyAsp Glu Pro Gly Arg Pro 565 570 575 Arg Ser Arg Gly Pro Val Gly Arg ThrGlu Pro Gly Glu Gly Pro Gln 580 585 590 Gln Glu Ile Lys Arg Arg Asp GlnPhe Pro Leu Thr Arg Ser Arg Ala 595 600 605 Ile Gln Glu Cys Arg Ser ProVal Pro Pro Tyr Thr Ala Asp Pro Pro 610 615 620 Glu Ser Arg Thr Lys AlaPro Ser Gly Arg Lys Arg Glu Pro Pro Ala 625 630 635 640 Gln Ala Val ArgPhe Leu Pro Trp Ala Thr Pro Gly Val Glu Asp Ser 645 650 655 Val Leu ProGln Thr Leu Glu Lys Asn Arg Ala Gly Pro Glu Ala Glu 660 665 670 Lys ArgLeu Arg Arg Gly Pro Glu Glu Asp Gly Pro Trp Gly Pro Trp 675 680 685 AspArg Arg Gly Thr Arg Ser Gln Gly Lys Gly Arg Arg Ala Arg Pro 690 695 700Thr Ser Pro Glu Leu Glu Ser Ser Asp Asp Ser Tyr Val Ser Ala Gly 705 710715 720 Glu Glu Pro Leu Glu Ala Pro Val Phe Glu Ile Pro Leu Gln Asn Met725 730 735 Val Val Ala Pro Gly Ala Asp Val Leu Leu Lys Cys Ile Ile ThrAla 740 745 750 Asn Pro Pro Pro Gln Val Ser Trp Lys Lys Asp Gly Ser MetLeu His 755 760 765 Ser Glu Gly Arg Leu Leu Ile Arg Ala Glu Gly Glu ArgHis Thr Leu 770 775 780 Leu Leu Arg Glu Ala Gln Ala Ala Asp Ala Gly SerTyr Thr Ala Thr 785 790 795 800 Ala Thr Asn Glu Leu Gly Gln Ala Thr CysAla Ser Ser Leu Ala Val 805 810 815 Arg Pro Gly Gly Ser Thr Ser Pro PheSer Ser Pro Ile Thr Ser Asp 820 825 830 Glu Glu Tyr Leu Ser Pro Pro GluGlu Phe Pro Glu Pro Gly Glu Thr 835 840 845 Trp Pro Arg Thr Pro Thr MetLys Leu Ser Pro Ser Gln Asp His Asp 850 855 860 Ser Ser Asp Ser Ser SerLys Ala Pro Pro Thr Phe Lys Val Ser Leu 865 870 875 880 Met Asp Gln SerVal Arg Glu Gly Gln Asp Val Ile Met Ser Ile Arg 885 890 895 Val Gln GlyGlu Pro Lys Pro Val Val Ser Trp Leu Arg Asn Arg Gln 900 905 910 Pro ValArg Pro Asp Gln Arg Arg Phe Ala Glu Glu Ala Glu Gly Gly 915 920 925 LeuCys Arg Leu Arg Ile Leu Ala Ala Glu Arg Gly Asp Ala Gly Phe 930 935 940Tyr Thr Cys Lys Ala Val Asn Glu Tyr Gly Ala Arg Gln Cys Glu Ala 945 950955 960 Arg Leu Glu Val Arg Ala His Pro Glu Ser Arg Ser Leu Ala Val Leu965 970 975 Ala Pro Leu Gln Asp Val Asp Val Gly Ala Gly Glu Met Ala LeuPhe 980 985 990 Glu Cys Leu Val Ala Gly Pro Ala Asp Val Glu Val Asp TrpLeu Cys 995 1000 1005 Arg Gly Arg Leu Leu Gln Pro Ala Leu Leu Lys CysLys Met His Phe 1010 1015 1020 Asp Gly Arg Lys Cys Lys Leu Leu Leu ThrSer Val His Glu Asp Asp 1025 1030 1035 1040 Ser Gly Val Tyr Thr Cys LysLeu Ser Thr Ala Lys Asp Glu Leu Thr 1045 1050 1055 Cys Ser Ala Arg LeuThr Val Arg Pro Ser Leu Ala Pro Leu Phe Thr 1060 1065 1070 Arg Leu LeuGlu Asp Val Glu Val Leu Glu Gly Arg Ala Ala Arg Leu 1075 1080 1085 AspCys Lys Ile Ser Gly Thr Pro Pro Pro Ser Val Thr Trp Thr His 1090 10951100 Phe Gly His Pro Val Asn Glu Gly Asp Asn Leu Arg Leu Arg Gln Asp1105 1110 1115 1120 Gly Gly Leu His Ser Leu His Ile Ala Arg Val Gly SerGlu Asp Glu 1125 1130 1135 Gly Leu Tyr Glu Val Ser Ala Thr Asn Thr HisGly Gln Ala His Cys 1140 1145 1150 Ser Ala Gln Leu Tyr Val Glu Glu ProArg Thr Ala Ala Ser Gly Pro 1155 1160 1165 Ser Ser Lys Leu Glu Lys MetPro Ser Ile Pro Glu Glu Pro Glu His 1170 1175 1180 Gly Asp Leu Glu ArgLeu Ser Ile Pro Asp Phe Leu Arg Pro Leu Gln 1185 1190 1195 1200 Asp LeuGlu Val Gly Leu Ala Lys Glu Ala Met Leu Glu Cys Gln Val 1205 1210 1215Thr Gly Leu Pro Tyr Pro Thr Ile Ser Trp Phe His Asn Gly His Arg 12201225 1230 Ile Gln Ser Ser Asp Asp Arg Arg Met Thr Gln Tyr Arg Asp IleHis 1235 1240 1245 Arg Leu Val Phe Pro Ala Val Gly Pro Gln His Ala GlyVal Tyr Lys 1250 1255 1260 Ser Val Ile Ala Asn Lys Leu Gly Lys Ala AlaCys Tyr Ala His Leu 1265 1270 1275 1280 Tyr Val Thr Asp Val Val Pro GlyPro Pro Asp Gly Ala Pro Glu Val 1285 1290 1295 Val Ala Val Thr Gly ArgMet Val Thr Leu Ser Trp Asn Pro Pro Arg 1300 1305 1310 Ser Leu Asp MetAla Ile Asp Pro Asp Ser Leu Thr Tyr Thr Val Gln 1315 1320 1325 His GlnVal Leu Gly Ser Asp Gln Trp Thr Ala Leu Val Thr Gly Leu 1330 1335 1340Arg Glu Pro Ala Trp Ala Ala Thr Gly Leu Lys Lys Gly Ile Gln His 13451350 1355 1360 Ile Phe Arg Val Leu Ser Ser Ser Gly Lys Ser Ser Ser LysPro Ser 1365 1370 1375 Ala Pro Ser Glu Pro Val Gln Leu Leu Glu His GlyPro Pro Leu Glu 1380 1385 1390 Glu Ala Pro Ala Val Leu Asp Lys Gln AspIle Val Tyr Val Val Glu 1395 1400 1405 Gly Gln Pro Ala Cys Val Thr ValThr Phe Asn His Val Glu Ala Gln 1410 1415 1420 Val Val Trp Arg Ser CysArg Gly Ala Leu Leu Glu Ala Arg Thr Gly 1425 1430 1435 1440 Val Tyr GluLeu Ser Gln Pro Asp Asp Asp Gln Tyr Cys Leu Arg Ile 1445 1450 1455 CysArg Val Ser Arg Arg Asp Leu Gly Pro Leu Thr Cys Ser Ala Arg 1460 14651470 Asn Arg His Gly Thr Lys Ala Cys Ser Val Thr Leu Glu Leu Ala Glu1475 1480 1485 Ala Pro Arg Phe Glu Ser Ile Met Glu Asp Val Glu Val GlyPro Gly 1490 1495 1500 Glu Thr Ala Arg Phe Ala Val Val Val Glu Gly LysPro Leu Pro Asp 1505 1510 1515 1520 Ile Met Trp Tyr Lys Asp Glu Val LeuLeu Ala Glu Ser Asn His Val 1525 1530 1535 Ser Phe Val Tyr Glu Glu AsnGlu Cys Ser Leu Val Leu Leu Ser Ala 1540 1545 1550 Gly Ser Gln Asp GlyGly Val Tyr Thr Cys Thr Ala Arg Asn Leu Ala 1555 1560 1565 Gly Glu ValSer Cys Lys Ala Glu Leu Ser Val Leu Ser Ala Gln Thr 1570 1575 1580 AlaMet Glu Val Glu Gly Val Gly Glu Asp Glu Glu His Arg Gly Arg 1585 15901595 1600 Arg Leu Ser Asp Tyr Tyr Asp Ile His Gln Glu Ile Gly Arg GlyAla 1605 1610 1615 Phe Ser Tyr Leu Arg Arg Val Val Glu Arg Ser Ser GlyLeu Glu Phe 1620 1625 1630 Ala Ala Lys Phe Ile Pro Ser Gln Ala Lys ProLys Ala Ser Ala Arg 1635 1640 1645 Arg Glu Ala Arg Leu Leu Ala Arg LeuGln His Gly Cys Val Leu Tyr 1650 1655 1660 Phe His Glu Ala Phe Glu ArgArg Arg Gly Leu Val Ile Val Thr Glu 1665 1670 1675 1680 Leu Cys Thr GluGlu Leu Leu Glu Arg Met Ala Arg Lys Pro Thr Val 1685 1690 1695 Cys GluSer Glu Thr Arg Thr Tyr Met Arg Gln Val Leu Glu Gly Ile 1700 1705 1710Cys Tyr Leu His Gln Ser His Val Leu His Leu Asp Val Lys Pro Glu 17151720 1725 Asn Leu Leu Val Trp Asp Gly Ala Gly Gly Glu Glu Gln Val ArgIle 1730 1735 1740 Cys Asp Phe Gly Asn Ala Gln Glu Leu Thr Pro Gly GluPro Gln Tyr 1745 1750 1755 1760 Cys Gln Tyr Gly Thr Pro Glu Phe Val AlaPro Glu Ile Val Asn Gln 1765 1770 1775 Ser Pro Val Ser Gly Val Thr AspIle Trp Pro Val Gly Val Val Ala 1780 1785 1790 Phe Leu Cys Leu Thr GlyIle Ser Pro Phe Val Gly Glu Asn Asp Arg 1795 1800 1805 Thr Thr Leu MetAsn Ile Arg Asn Tyr Asn Val Ala Phe Glu Glu Thr 1810 1815 1820 Thr PheLeu Ser Leu Ser Arg Glu Ala Arg Gly Phe Leu Ile Lys Val 1825 1830 18351840 Leu Val Gln Asp Arg Leu Arg Pro Thr Ala Glu Glu Thr Leu Glu His1845 1850 1855 Pro Trp Phe Lys Thr Glu Ala Lys Gly Ala Glu Val Ser ThrAsp His 1860 1865 1870 Leu Lys Leu Phe Leu Ser Arg Arg Arg Trp Gln ArgSer Gln Ile Ser 1875 1880 1885 Tyr Lys Cys His Leu Val Leu Arg Pro IlePro Glu Leu Leu Arg Ala 1890 1895 1900 Pro Pro Glu Arg Val Trp Val AlaMet Pro Arg Arg Gln Pro Pro Ser 1905 1910 1915 1920 Gly Gly Leu Ser SerSer Ser Asp Ser Glu Glu Glu Glu Leu Glu Glu 1925 1930 1935 Leu Pro SerVal Pro Arg Pro Leu Gln Pro Glu Phe Ser Gly Ser Arg 1940 1945 1950 ValSer Leu Thr Asp Ile Pro Thr Glu Asp Glu Ala Leu Gly Thr Pro 1955 19601965 Glu Ala Gly Ala Ala Thr Pro Met Asp Trp Gln Glu Gln Glu Arg Thr1970 1975 1980 Pro Ser Lys Asp Gln Glu Ala Pro Ser Pro Glu Ala Leu ProSer Pro 1985 1990 1995 2000 Gly Gln Glu Ser Pro Asp Gly Pro Ser Pro ArgArg Pro Glu Leu Arg 2005 2010 2015 Arg Gly Ser Ser Ala Glu Ser Ala LeuPro Arg Val Gly Ser Arg Glu 2020 2025 2030 Pro Gly Arg Ser Leu His LysAla Ala Ser Val Glu Leu Pro Gln Arg 2035 2040 2045 Arg Ser Pro Ser ProGly Ala Thr Arg Leu Thr Arg Gly Gly Leu Gly 2050 2055 2060 Glu Gly GluTyr Ala Gln Arg Leu Gln Ala Leu Arg Gln Arg Leu Leu 2065 2070 2075 2080Arg Gly Gly Pro Glu Asp Gly Lys Val Ser Gly Leu Arg Gly Pro Leu 20852090 2095 Leu Glu Ser Leu Gly Gly Arg Ala Arg Asp Pro Arg Met Ala ArgAla 2100 2105 2110 Ala Ser Ser Glu Ala Ala Pro His His Gln Pro Pro ProGlu Ser Arg 2115 2120 2125 Gly Leu Gln Lys Ser Ser Ser Phe Ser Gln GlyGlu Ala Glu Pro Arg 2130 2135 2140 Gly Arg His Arg Arg Ala Gly Ala ProLeu Glu Ile Pro Val Ala Arg 2145 2150 2155 2160 Leu Gly Ala Arg Arg LeuGln Glu Ser Pro Ser Leu Ser Ala Leu Ser 2165 2170 2175 Glu Thr Gln ProPro Ser Pro Ala Arg Pro Ser Val Pro Lys Leu Ser 2180 2185 2190 Ile ThrLys Ser Pro Glu Pro Ser Ala Val Thr Ser Arg Asp Ser Pro 2195 2200 2205Gln Pro Pro Glu Pro Gln Pro Val Pro Glu Lys Val Pro Glu Pro Lys 22102215 2220 Pro Glu Pro Val Arg Ala Ala Lys Pro Ala Gln Pro Pro Leu AlaLeu 2225 2230 2235 2240 Gln Met Pro Thr Gln Pro Leu Thr Pro Tyr Ala GlnIle Met Gln Ser 2245 2250 2255 Leu Gln Leu Ser Ser Pro Thr Leu Ser ProGln Asp Pro Ala Val Pro 2260 2265 2270 Pro Ser Glu Pro Lys Pro His AlaAla Val Phe Ala Arg Val Ala Ser 2275 2280 2285 Pro Pro Pro Gly Val SerGlu Lys Arg Val Pro Ser Ala Arg Thr Pro 2290 2295 2300 Pro Val Leu AlaGlu Lys Ala Arg Val Pro Thr Val Pro Pro Arg Pro 2305 2310 2315 2320 GlySer Ser Leu Ser Gly Ser Ile Glu Asn Leu Glu Ser Glu Ala Val 2325 23302335 Phe Glu Ala Lys Phe Lys Arg Ser Arg Glu Ser Pro Leu Ser Arg Gly2340 2345 2350 Leu Arg Leu Leu Ser Arg Ser Arg Ser Glu Glu Arg Gly ProPhe Arg 2355 2360 2365 Gly Ala Glu Asp Asp Gly Ile Tyr Arg Pro Ser ProAla Gly Thr Pro 2370 2375 2380 Leu Glu Leu Val Arg Arg Pro Glu Arg SerArg Ser Val Gln Asp Leu 2385 2390 2395 2400 Arg Val Ala Gly Glu Pro GlyLeu Val Arg Arg Leu Ser Leu Ser Leu 2405 2410 2415 Ser Gln Lys Leu ArgArg Thr Pro Pro Gly Gln Arg His Pro Ala Trp 2420 2425 2430 Glu Ser ArgSer Gly Asp Gly Glu Ser Ser Glu Gly Gly Ser Ser Ala 2435 2440 2445 ArgAla Ser Pro Val Leu Ala Val Arg Arg Arg Leu Ser Ser Thr Leu 2450 24552460 Glu Arg Leu Ser Ser Arg Leu Gln Arg Ser Gly Ser Ser Glu Asp Ser2465 2470 2475 2480 Gly Gly Ala Ser Gly Arg Ser Thr Pro Leu Phe Gly ArgLeu Arg Arg 2485 2490 2495 Ala Thr Ser Glu Gly Glu Ser Leu Arg Arg LeuGly Val Pro His Asn 2500 2505 2510 Gln Leu Gly Ser Gln Thr Gly Ala ThrThr Pro Ser Ala Glu Ser Leu 2515 2520 2525 Gly Ser Glu Ala Ser Gly ThrSer Gly Ser Ser Ala Pro Gly Glu Ser 2530 2535 2540 Arg Ser Arg His ArgTrp Gly Leu Ser Arg Leu Arg Lys Asp Lys Gly 2545 2550 2555 2560 Leu SerGln Pro Asn Leu Ser Ser Ser Val Gln Glu Asp Leu Gly His 2565 2570 2575Gln Tyr Val Pro Ser Glu Ser Asp Phe Pro Pro Val Phe His Ile Lys 25802585 2590 Leu Lys Asp Gln Val Leu Leu Glu Gly Glu Ala Ala Thr Leu LeuCys 2595 2600 2605 Leu Pro Ala Ala Cys Pro Ala Pro Arg Ile Ser Trp MetLys Asp Lys 2610 2615 2620 Gln Ser Leu Arg Ser Glu Pro Ser Val Val IleVal Ser Cys Lys Asp 2625 2630 2635 2640 Gly Arg Gln Leu Leu Ser Ile ProArg Ala Gly Lys Arg His Ala Gly 2645 2650 2655 Leu Tyr Glu Cys Ser AlaThr Asn Val Leu Gly Ser Ile Thr Ser Ser 2660 2665 2670 Cys Thr Val AlaVal Ala Arg Ile Pro Gly Lys Leu Ala Pro Pro Glu 2675 2680 2685 Val ProGln Thr Tyr His Asp Thr Ala Leu Val Val Trp Lys Pro Gly 2690 2695 2700Asp Gly Arg Ala Pro Cys Thr Tyr Thr Leu Glu Arg Arg Val Asp Gly 27052710 2715 2720 Glu Ser Val Trp His Pro Val Ser Ser Gly Ile Pro Asp CysTyr Tyr 2725 2730 2735 Asn Val Thr Gln Leu Pro Val Gly Val Thr Val ArgPhe Arg Val Ala 2740 2745 2750 Cys Ser Asn Arg Ala Gly Gln Gly Pro PheSer Asn Pro Ser Glu Lys 2755 2760 2765 Val Phe Ile Arg Gly Thr Pro AspSer Pro Ala Gln Pro Ala Ala Ala 2770 2775 2780 Pro Arg Asp Ala Pro ValThr Ser Gly Pro Thr Arg Ala Pro Pro Pro 2785 2790 2795 2800 Asp Ser ProThr Ser Leu Ala Pro Thr Pro Ala Leu Ala Pro Pro Ala 2805 2810 2815 SerGln Ala Ser Thr Leu Ser Pro Ser Thr Ser Ser Met Ser Ala Asn 2820 28252830 Gln Ala Leu Ser Ser Leu Lys Ala Val Gly Pro Pro Pro Ala Thr Pro2835 2840 2845 Pro Arg Lys His Arg Gly Leu Leu Ala Thr Gln Gln Ala GluPro Ser 2850 2855 2860 Pro Pro Ser Ile Val Val Thr Pro Ser Glu Pro ArgSer Phe Val Pro 2865 2870 2875 2880 Asp Thr Gly Thr Leu Thr Pro Thr SerSer Pro Gln Gly Val Lys Pro 2885 2890 2895 Ala Pro Ser Ser Thr Ser LeuTyr Met Val Thr Ser Phe Val Ser Ala 2900 2905 2910 Pro Pro Ala Pro GlnAla Pro Ala Pro Glu Pro Pro Pro Glu Pro Thr 2915 2920 2925 Lys Val ThrVal Arg Ser Leu Ser Pro Ala Lys Glu Val Val Ser Ser 2930 2935 2940 ProThr Pro Glu Ser Thr Thr Leu Arg Gln Gly Pro Leu Arg Asn Pro 2945 29502955 2960 Thr Pro Ser Trp Arg Arg Arg Pro Gly Gly Ala Leu Ala Leu CysGly 2965 2970 2975 His Ala Gly Arg Met Leu Arg Ala Glu Arg Leu Ser ProArg Phe Val 2980 2985 2990 Pro Tyr Ala Ala Glu Gly Lys Arg Arg Val LeuGln Glu Tyr Glu Val 2995 3000 3005 Leu Arg Thr Leu His His Glu Arg LeuMet Ser Leu His Glu Ala Tyr 3010 3015 3020 Ile Thr Pro Arg Tyr Leu ValLeu Ile Ala Glu Ser Cys Gly Asn Arg 3025 3030 3035 3040 Glu Leu Leu CysGly Leu Ser Asp Arg Phe Arg Tyr Ser Glu Asp Asp 3045 3050 3055 Val AlaThr Tyr Val Val Gln Leu Leu Gln Gly Leu Asp Tyr Leu His 3060 3065 3070Gly His His Val Leu His Leu Asp Ile Lys Pro Asp Asn Leu Leu Leu 30753080 3085 Ala Ala Asp Asn Ala Leu Lys Ile Val Asp Phe Gly Ser Ala GlnPro 3090 3095 3100 Tyr Asn Pro Gln Ala Leu Lys Pro Leu Gly His Arg ThrGly Thr Leu 3105 3110 3115 3120 Glu Phe Met Ala Pro Glu Met Val Lys GlyAsp Pro Ile Gly Ser Ala 3125 3130 3135 Thr Asp Ile Trp Gly Ala Gly ValLeu Thr Tyr Ile Met Leu Ser Gly 3140 3145 3150 Tyr Ser Pro Phe Tyr GluPro Asp Pro Gln Glu Thr Glu Ala Arg Ile 3155 3160 3165 Val Gly Gly ArgPhe Asp Ala Phe Gln Leu Tyr Pro Asn Thr Ser Gln 3170 3175 3180 Ser AlaThr Leu Phe Leu Arg Lys Val Leu Ser Val His Pro Trp Ser 3185 3190 31953200 Arg Pro Ser Leu Gln Asp Cys Leu Ala His Pro Trp Leu Gln Asp Ala3205 3210 3215 Tyr Leu Met Lys Leu Arg Arg Gln Thr Leu Thr Phe Thr ThrAsn Arg 3220 3225 3230 Leu Lys Glu Phe Leu Gly Glu Gln Arg Arg Arg ArgAla Glu Ala Ala 3235 3240 3245 Thr Arg His Lys Val Leu Leu Arg Ser TyrPro Gly Ser Pro 3250 3255 3260 5 2231 PRT Homo sapiens 5 Gly Glu Met AlaLeu Phe Glu Cys Leu Val Ala Gly Pro Thr Asp Val 1 5 10 15 Glu Val AspTrp Leu Cys Arg Gly Arg Leu Leu Gln Pro Ala Leu Leu 20 25 30 Lys Cys LysMet His Phe Asp Gly Arg Lys Cys Lys Leu Leu Leu Thr 35 40 45 Ser Val HisGlu Asp Asp Ser Gly Val Tyr Thr Cys Lys Leu Ser Thr 50 55 60 Ala Lys AspGlu Leu Thr Cys Ser Ala Arg Leu Thr Val Arg Pro Ser 65 70 75 80 Leu AlaPro Leu Phe Thr Arg Leu Leu Glu Asp Val Glu Val Leu Glu 85 90 95 Gly ArgAla Ala Arg Phe Asp Cys Lys Ile Ser Gly Thr Pro Pro Pro 100 105 110 ValVal Thr Trp Thr His Phe Gly Cys Pro Met Glu Glu Ser Glu Asn 115 120 125Leu Arg Leu Arg Gln Asp Gly Gly Leu His Ser Leu His Ile Ala His 130 135140 Val Gly Ser Glu Asp Glu Gly Leu Tyr Ala Val Ser Ala Val Asn Thr 145150 155 160 His Gly Gln Ala His Cys Ser Ala Gln Leu Tyr Val Glu Glu ProArg 165 170 175 Thr Ala Ala Ser Gly Pro Ser Ser Lys Leu Glu Lys Met ProSer Ile 180 185 190 Pro Glu Glu Pro Glu Gln Gly Glu Leu Glu Arg Leu SerIle Pro Asp 195 200 205 Phe Leu Arg Pro Leu Gln Asp Leu Glu Val Gly LeuAla Lys Glu Ala 210 215 220 Met Leu Glu Cys Gln Val Thr Gly Leu Pro TyrPro Thr Ile Ser Trp 225 230 235 240 Phe His Asn Gly His Arg Ile Gln SerSer Asp Asp Arg Arg Met Thr 245 250 255 Gln Tyr Arg Asp Val His Arg LeuVal Phe Pro Ala Val Gly Pro Gln 260 265 270 His Ala Gly Val Tyr Lys SerVal Ile Ala Asn Lys Leu Gly Lys Ala 275 280 285 Ala Cys Tyr Ala His LeuTyr Val Thr Asp Val Val Pro Gly Pro Pro 290 295 300 Asp Gly Ala Pro GlnVal Val Ala Val Thr Gly Arg Met Val Thr Leu 305 310 315 320 Thr Trp AsnPro Pro Arg Ser Leu Asp Met Ala Ile Asp Pro Asp Ser 325 330 335 Leu ThrTyr Thr Val Gln His Gln Val Leu Gly Ser Asp Gln Trp Thr 340 345 350 AlaLeu Val Thr Gly Leu Arg Glu Pro Gly Trp Ala Ala Thr Gly Leu 355 360 365Arg Lys Gly Val Gln His Ile Phe Arg Val Leu Ser Thr Thr Val Lys 370 375380 Ser Ser Ser Lys Pro Ser Pro Pro Ser Glu Pro Val Gln Leu Leu Glu 385390 395 400 His Gly Pro Thr Leu Glu Glu Ala Pro Ala Met Leu Asp Lys ProAsp 405 410 415 Ile Val Tyr Val Val Glu Gly Gln Pro Ala Ser Val Thr ValThr Phe 420 425 430 Asn His Val Glu Ala Gln Val Val Trp Arg Ser Cys ArgGly Ala Leu 435 440 445 Leu Glu Ala Arg Ala Gly Val Tyr Glu Leu Ser GlnPro Asp Asp Asp 450 455 460 Gln Tyr Cys Leu Arg Ile Cys Arg Val Ser ArgArg Asp Met Gly Ala 465 470 475 480 Leu Thr Cys Thr Ala Arg Asn Arg HisGly Thr Gln Thr Cys Ser Val 485 490 495 Thr Leu Glu Leu Ala Glu Ala ProArg Phe Glu Ser Ile Met Glu Asp 500 505 510 Val Glu Val Gly Ala Gly GluThr Ala Arg Phe Ala Val Val Val Glu 515 520 525 Gly Lys Pro Leu Pro AspIle Met Trp Tyr Lys Asp Glu Val Leu Leu 530 535 540 Thr Glu Ser Ser HisVal Ser Phe Val Tyr Glu Glu Asn Glu Cys Ser 545 550 555 560 Leu Val ValLeu Ser Thr Gly Ala Gln Asp Gly Gly Val Tyr Thr Cys 565 570 575 Thr AlaGln Asn Leu Ala Gly Glu Val Ser Cys Lys Ala Glu Leu Ala 580 585 590 ValHis Ser Ala Gln Thr Ala Met Glu Val Glu Gly Val Gly Glu Asp 595 600 605Glu Asp His Arg Gly Arg Arg Leu Ser Asp Phe Tyr Asp Ile His Gln 610 615620 Glu Ile Gly Arg Gly Ala Phe Ser Tyr Leu Arg Arg Ile Val Glu Arg 625630 635 640 Ser Ser Gly Leu Glu Phe Ala Ala Lys Phe Ile Pro Ser Gln AlaLys 645 650 655 Pro Lys Ala Ser Ala Arg Arg Glu Ala Arg Leu Leu Ala ArgLeu Gln 660 665 670 His Asp Cys Val Leu Tyr Phe His Glu Ala Phe Glu ArgArg Arg Gly 675 680 685 Leu Val Ile Val Thr Glu Leu Cys Thr Glu Glu LeuLeu Glu Arg Ile 690 695 700 Ala Arg Lys Pro Thr Val Cys Glu Ser Glu IleArg Ala Tyr Met Arg 705 710 715 720 Gln Val Leu Glu Gly Ile His Tyr LeuHis Gln Ser His Val Leu His 725 730 735 Leu Asp Val Lys Pro Glu Asn LeuLeu Val Trp Asp Gly Ala Ala Gly 740 745 750 Glu Gln Gln Val Arg Ile CysAsp Phe Gly Asn Ala Gln Glu Leu Thr 755 760 765 Pro Gly Glu Pro Gln TyrCys Gln Tyr Gly Thr Pro Glu Phe Val Ala 770 775 780 Pro Glu Ile Val AsnGln Ser Pro Val Ser Gly Val Thr Asp Ile Trp 785 790 795 800 Pro Val GlyVal Val Ala Phe Leu Cys Leu Thr Gly Ile Ser Pro Phe 805 810 815 Val GlyGlu Asn Asp Arg Thr Thr Leu Met Asn Ile Arg Asn Tyr Asn 820 825 830 ValAla Phe Glu Glu Thr Thr Phe Leu Ser Leu Ser Arg Glu Ala Arg 835 840 845Gly Phe Leu Ile Lys Val Leu Val Gln Asp Arg Leu Arg Pro Thr Ala 850 855860 Glu Glu Thr Leu Glu His Pro Trp Phe Lys Thr Gln Ala Lys Gly Ala 865870 875 880 Glu Val Ser Thr Asp His Leu Lys Leu Phe Leu Ser Arg Arg ArgTrp 885 890 895 Gln Arg Ser Gln Ile Ser Tyr Lys Cys His Leu Val Leu ArgPro Ile 900 905 910 Pro Glu Leu Leu Arg Ala Pro Pro Glu Arg Val Trp ValThr Met Pro 915 920 925 Arg Arg Pro Pro Pro Ser Gly Gly Leu Ser Ser SerSer Asp Ser Glu 930 935 940 Glu Glu Glu Leu Glu Glu Leu Pro Ser Val ProArg Pro Leu Gln Pro 945 950 955 960 Glu Phe Ser Gly Ser Arg Val Ser LeuThr Asp Ile Pro Thr Glu Asp 965 970 975 Glu Ala Leu Gly Thr Pro Glu ThrGly Ala Ala Thr Pro Met Asp Trp 980 985 990 Gln Glu Gln Gly Arg Ala ProSer Gln Asp Gln Glu Ala Pro Ser Pro 995 1000 1005 Glu Ala Leu Pro SerPro Gly Gln Glu Pro Ala Ala Gly Ala Ser Pro 1010 1015 1020 Arg Arg GlyGlu Leu Arg Arg Gly Ser Ser Ala Glu Ser Ala Leu Pro 1025 1030 1035 1040Arg Ala Gly Pro Arg Glu Leu Gly Arg Gly Leu His Lys Ala Ala Ser 10451050 1055 Val Glu Leu Pro Gln Arg Arg Ser Pro Gly Pro Gly Ala Thr ArgLeu 1060 1065 1070 Ala Arg Gly Gly Leu Gly Glu Gly Glu Tyr Ala Gln ArgLeu Gln Ala 1075 1080 1085 Leu Arg Gln Arg Leu Leu Arg Gly Gly Pro GluAsp Gly Lys Val Ser 1090 1095 1100 Gly Leu Arg Gly Pro Leu Leu Glu SerLeu Gly Gly Arg Ala Arg Asp 1105 1110 1115 1120 Pro Arg Met Ala Arg AlaAla Ser Ser Glu Ala Ala Pro His His Gln 1125 1130 1135 Pro Pro Leu GluAsn Arg Gly Leu Gln Lys Ser Ser Ser Phe Ser Gln 1140 1145 1150 Gly GluAla Glu Pro Arg Gly Arg His Arg Arg Ala Gly Ala Pro Leu 1155 1160 1165Glu Ile Pro Val Ala Arg Leu Gly Ala Arg Arg Leu Gln Glu Ser Pro 11701175 1180 Ser Leu Ser Ala Leu Ser Glu Ala Gln Pro Ser Ser Pro Ala ArgPro 1185 1190 1195 1200 Ser Ala Pro Lys Pro Ser Thr Pro Lys Ser Ala GluPro Ser Ala Thr 1205 1210 1215 Thr Pro Ser Asp Ala Pro Gln Pro Pro AlaPro Gln Pro Ala Gln Asp 1220 1225 1230 Lys Ala Pro Glu Pro Arg Pro GluPro Val Arg Ala Ser Lys Pro Ala 1235 1240 1245 Pro Pro Pro Gln Ala LeuGln Thr Leu Ala Leu Pro Leu Thr Pro Tyr 1250 1255 1260 Ala Gln Ile IleGln Ser Leu Gln Leu Ser Gly His Ala Gln Gly Pro 1265 1270 1275 1280 SerGln Gly Pro Ala Ala Pro Pro Ser Glu Pro Lys Pro His Ala Ala 1285 12901295 Val Phe Ala Arg Val Ala Ser Pro Pro Pro Gly Ala Pro Glu Lys Arg1300 1305 1310 Val Pro Ser Ala Gly Gly Pro Pro Val Leu Ala Glu Lys AlaArg Val 1315 1320 1325 Pro Thr Val Pro Pro Arg Pro Gly Ser Ser Leu SerSer Ser Ile Glu 1330 1335 1340 Asn Leu Glu Ser Glu Ala Val Phe Glu AlaLys Phe Lys Arg Ser Arg 1345 1350 1355 1360 Glu Ser Pro Leu Ser Leu GlyLeu Arg Leu Leu Ser Arg Ser Arg Ser 1365 1370 1375 Glu Glu Arg Gly ProPhe Arg Gly Ala Glu Glu Glu Asp Gly Ile Tyr 1380 1385 1390 Arg Pro SerPro Ala Gly Thr Pro Leu Glu Leu Val Arg Arg Pro Glu 1395 1400 1405 ArgSer Arg Ser Val Gln Asp Leu Arg Ala Val Gly Glu Pro Gly Leu 1410 14151420 Val Arg Arg Leu Ser Leu Ser Leu Ser Gln Arg Leu Arg Arg Thr Pro1425 1430 1435 1440 Pro Ala Gln Arg His Pro Ala Trp Glu Ala Arg Gly GlyAsp Gly Glu 1445 1450 1455 Ser Ser Glu Gly Gly Ser Ser Ala Arg Gly SerPro Val Leu Ala Met 1460 1465 1470 Arg Arg Arg Leu Ser Phe Thr Leu GluArg Leu Ser Ser Arg Leu Gln 1475 1480 1485 Arg Ser Gly Ser Ser Glu AspSer Gly Gly Ala Ser Gly Arg Ser Thr 1490 1495 1500 Pro Leu Phe Gly ArgLeu Arg Arg Ala Thr Ser Glu Gly Glu Ser Leu 1505 1510 1515 1520 Arg ArgLeu Gly Leu Pro His Asn Gln Leu Ala Ala Gln Ala Gly Ala 1525 1530 1535Thr Thr Pro Ser Ala Glu Ser Leu Gly Ser Glu Ala Ser Ala Thr Ser 15401545 1550 Gly Ser Ser Ala Pro Gly Glu Ser Arg Ser Arg Leu Arg Trp GlyPhe 1555 1560 1565 Ser Arg Pro Arg Lys Asp Lys Gly Leu Ser Pro Pro AsnLeu Ser Ala 1570 1575 1580 Ser Val Gln Glu Glu Leu Gly His Gln Tyr ValArg Ser Glu Ser Asp 1585 1590 1595 1600 Phe Pro Pro Val Phe His Ile LysLeu Lys Asp Gln Val Leu Leu Glu 1605 1610 1615 Gly Glu Ala Ala Thr LeuLeu Cys Leu Pro Ala Ala Cys Pro Ala Pro 1620 1625 1630 His Ile Ser TrpMet Lys Asp Lys Lys Ser Leu Arg Ser Glu Pro Ser 1635 1640 1645 Val IleIle Val Ser Cys Lys Asp Gly Arg Gln Leu Leu Ser Ile Pro 1650 1655 1660Arg Ala Gly Lys Arg His Ala Gly Leu Tyr Glu Cys Ser Ala Thr Asn 16651670 1675 1680 Val Leu Gly Ser Ile Thr Ser Ser Cys Thr Val Ala Val AlaArg Val 1685 1690 1695 Pro Gly Lys Leu Ala Pro Pro Glu Val Thr Gln ThrTyr Gln Asp Thr 1700 1705 1710 Ala Leu Val Leu Trp Lys Pro Gly Asp SerArg Ala Pro Cys Thr Tyr 1715 1720 1725 Thr Leu Glu Arg Arg Val Asp GlyGlu Ser Val Trp His Pro Val Ser 1730 1735 1740 Ser Gly Ile Pro Asp CysTyr Tyr Asn Val Thr His Leu Pro Val Gly 1745 1750 1755 1760 Val Thr ValArg Phe Arg Val Ala Cys Ala Asn Arg Ala Gly Gln Gly 1765 1770 1775 ProPhe Ser Asn Ser Ser Glu Lys Val Phe Val Arg Gly Thr Gln Asp 1780 17851790 Ser Ser Ala Val Pro Ser Ala Ala His Gln Glu Ala Pro Val Thr Ser1795 1800 1805 Arg Pro Ala Arg Ala Arg Pro Pro Asp Ser Pro Thr Ser LeuAla Pro 1810 1815 1820 Pro Leu Ala Pro Ala Ala Pro Thr Pro Pro Ser ValThr Val Ser Pro 1825 1830 1835 1840 Ser Ser Pro Pro Thr Pro Pro Ser GlnAla Leu Ser Ser Leu Lys Ala 1845 1850 1855 Val Gly Pro Pro Pro Gln ThrPro Pro Arg Arg His Arg Gly Leu Gln 1860 1865 1870 Ala Ala Arg Pro AlaGlu Pro Thr Leu Pro Ser Thr His Val Thr Pro 1875 1880 1885 Ser Glu ProLys Pro Phe Val Leu Asp Thr Gly Thr Pro Ile Pro Ala 1890 1895 1900 SerThr Pro Gln Gly Val Lys Pro Val Ser Ser Ser Thr Pro Val Tyr 1905 19101915 1920 Val Val Thr Ser Phe Val Ser Ala Pro Pro Ala Pro Glu Pro ProAla 1925 1930 1935 Pro Glu Pro Pro Pro Glu Pro Thr Lys Val Thr Val GlnSer Leu Ser 1940 1945 1950 Pro Ala Lys Glu Val Val Ser Ser Pro Gly SerSer Pro Arg Ser Ser 1955 1960 1965 Pro Arg Pro Glu Gly Thr Thr Leu ArgGln Gly Pro Pro Gln Lys Pro 1970 1975 1980 Tyr Thr Phe Leu Glu Glu LysAla Arg Gly Arg Phe Gly Val Val Arg 1985 1990 1995 2000 Ala Cys Arg GluAsn Ala Thr Gly Arg Thr Phe Val Ala Lys Ile Val 2005 2010 2015 Pro TyrAla Ala Glu Gly Lys Pro Arg Val Leu Gln Glu Tyr Glu Val 2020 2025 2030Leu Arg Thr Leu His His Glu Arg Ile Met Ser Leu His Glu Ala Tyr 20352040 2045 Ile Thr Pro Arg Tyr Leu Val Leu Ile Ala Glu Ser Cys Gly AsnArg 2050 2055 2060 Glu Leu Leu Cys Gly Leu Ser Asp Arg Phe Arg Tyr SerGlu Asp Asp 2065 2070 2075 2080 Val Ala Thr Tyr Met Val Gln Leu Leu GlnGly Leu Asp Tyr Leu His 2085 2090 2095 Gly His His Val Leu His Leu AspIle Lys Pro Asp Asn Leu Leu Leu 2100 2105 2110 Ala Pro Asp Asn Ala LeuLys Ile Val Asp Phe Gly Ser Ala Gln Pro 2115 2120 2125 Tyr Asn Pro GlnAla Leu Arg Pro Leu Gly His Arg Thr Gly Thr Leu 2130 2135 2140 Glu PheMet Ala Pro Glu Met Val Lys Gly Glu Pro Ile Gly Ser Ala 2145 2150 21552160 Thr Asp Ile Trp Gly Ala Gly Val Leu Thr Tyr Ile Met Leu Ser Gly2165 2170 2175 Arg Ser Pro Phe Tyr Glu Pro Asp Pro Gln Glu Thr Glu AlaArg Ile 2180 2185 2190 Val Gly Gly Arg Phe Asp Ala Phe Gln Leu Tyr ProAsn Thr Ser Gln 2195 2200 2205 Ser Ala Thr Leu Phe Leu Arg Lys Val LeuSer Val His Pro Trp Ser 2210 2215 2220 Arg Pro Ser Ser Cys Leu Ser 22252230

That which is claimed is:
 1. An isolated peptide consisting of an aminoacid sequence selected from the group consisting of: (a) an amino acidsequence shown in SEQ ID NO:2; (b) an amino acid sequence of an allelicvariant of an amino acid sequence shown in SEQ ID NO:2, wherein saidallelic variant is encoded by a nucleic acid molecule that hybridizesunder stringent conditions to the opposite strand of a nucleic acidmolecule shown in SEQ ID NOS:1 or 3; (c) an amino acid sequence of anortholog of an amino acid sequence shown in SEQ ID NO:2, wherein saidortholog is encoded by a nucleic acid molecule that hybridizes understringent conditions to the opposite strand of a nucleic acid moleculeshown in SEQ ID NOS:1 or 3; and (d) a fragment of an amino acid sequenceshown in SEQ ID NO:2, wherein said fragment comprises at least 10contiguous amino acids.
 2. An isolated peptide comprising an amino acidsequence selected from the group consisting of: (a) an amino acidsequence shown in SEQ ID NO:2; (b) an amino acid sequence of an allelicvariant of an amino acid sequence shown in SEQ ID NO:2, wherein saidallelic variant is encoded by a nucleic acid molecule that hybridizesunder stringent conditions to the opposite strand of a nucleic acidmolecule shown in SEQ ID NOS:1or 3; (c) an amino acid sequence of anortholog of an amino acid sequence shown in SEQ ID NO:2, wherein saidortholog is encoded by a nucleic acid molecule that hybridizes understringent conditions to the opposite strand of a nucleic acid moleculeshown in SEQ ID NOS:1 or 3; and (d) a fragment of an amino acid sequenceshown in SEQ ID NO:2, wherein said fragment comprises at least 10contiguous amino acids.
 3. An isolated antibody that selectively bindsto a peptide of claim
 2. 4. An isolated nucleic acid molecule consistingof a nucleotide sequence selected from the group consisting of: (a) anucleotide sequence that encodes an amino acid sequence shown in SEQ IDNO:2; (b) a nucleotide sequence that encodes of an allelic variant of anamino acid sequence shown in SEQ ID NO:2, wherein said nucleotidesequence hybridizes under stringent conditions to the opposite strand ofa nucleic acid molecule shown in SEQ ID NOS: 1 or 3; (c) a nucleotidesequence that encodes an ortholog of an amino acid sequence shown in SEQID NO:2, wherein said nucleotide sequence hybridizes under stringentconditions to the opposite strand of a nucleic acid molecule shown inSEQ ID NOS:1 or 3; (d) a nucleotide sequence that encodes a fragment ofan amino acid sequence shown in SEQ ID NO:2, wherein said fragmentcomprises at least 10 contiguous amino acids; and (e) a nucleotidesequence that is the complement of a nucleotide sequence of (a)-(d). 5.An isolated nucleic acid molecule comprising a nucleotide sequenceselected from the group consisting of: (a) a nucleotide sequence thatencodes an amino acid sequence shown in SEQ ID NO:2; (b) a nucleotidesequence that encodes of an allelic variant of an amino acid sequenceshown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes understringent conditions to the opposite strand of a nucleic acid moleculeshown in SEQ ID NOS:1 or 3; (c) a nucleotide sequence that encodes anortholog of an amino acid sequence shown in SEQ ID NO:2, wherein saidnucleotide sequence hybridizes under stringent conditions to theopposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3;(d) a nucleotide sequence that encodes a fragment of an amino acidsequence shown in SEQ ID NO:2, wherein said fragment comprises at least10 contiguous amino acids; and (e) a nucleotide sequence that is thecomplement of a nucleotide sequence of (a)-(d).
 6. A gene chipcomprising a nucleic acid molecule of claim
 5. 7. A transgenic non-humananimal comprising a nucleic acid molecule of claim
 5. 8. A nucleic acidvector comprising a nucleic acid molecule of claim
 5. 9. A host cellcontaining the vector of claim
 8. 10. A method for producing any of thepeptides of claim 1 comprising introducing a nucleotide sequenceencoding any of the amino acid sequences in (a)-(d) into a host cell,and culturing the host cell under conditions in which the peptides areexpressed from the nucleotide sequence.
 11. A method for producing anyof the peptides of claim 2 comprising introducing a nucleotide sequenceencoding any of the amino acid sequences in (a)-(d) into a host cell,and culturing the host cell under conditions in which the peptides areexpressed from the nucleotide sequence.
 12. A method for detecting thepresence of any of the peptides of claim 2 in a sample, said methodcomprising contacting said sample with a detection agent thatspecifically allows detection of the presence of the peptide in thesample and then detecting the presence of the peptide.
 13. A method fordetecting the presence of a nucleic acid molecule of claim
 5. in asample, said method comprising contacting the sample with anoligonucleotide that hybridizes to said nucleic acid molecule understringent conditions and determining whether the oligonucleotide bindsto said nucleic acid molecule in the sample.
 14. A method foridentifying a modulator of a peptide of claim 2, said method comprisingcontacting said peptide with an agent and determining if said agent hasmodulated the function or activity of said peptide.
 15. The method ofclaim 14, wherein said agent is administered to a host cell comprisingan expression vector that expresses said peptide.
 16. A method foridentifying an agent that binds to any of the peptides of claim 2, saidmethod comprising contacting the peptide with an agent and assaying thecontacted mixture to determine whether a complex is formed with theagent bound to the peptide.
 17. A pharmaceutical composition comprisingan agent identified by the method of claim 16 and a pharmaceuticallyacceptable carrier therefor.
 18. A method for treating a disease orcondition mediated by a human kinase protein, said method comprisingadministering to a patient a pharmaceutically effective amount of anagent identified by the method of claim
 16. 19. A method for identifyinga modulator of the expression of a peptide of claim 2, said methodcomprising contacting a cell expressing said peptide with an agent, anddetermining if said agent has modulated the expression of said peptide.20. An isolated human kinase peptide having an amino acid sequence thatshares at least 70% homology with an amino acid sequence shown in SEQ IDNO:2.
 21. A peptide according to claim 20 that shares at least 90percent homology with an amino acid sequence shown in SEQ ID NO:2. 22.An isolated nucleic acid molecule encoding a human kinase peptide, saidnucleic acid molecule sharing at least 80 percent homology with anucleic acid molecule shown in SEQ ID. NOS:1 or
 3. 23. A nucleic acidmolecule according to claim 22 that shares at least 90 percent homologywith a nucleic acid molecule shown in SEQ ID NOS:1 or 3.